RU2810094C2 - Procoagulant antibodies - Google Patents

Abstract

FIELD: biochemistry.

SUBSTANCE: invention relates to a procoagulant bispecific antibody that specifically binds FIX and FX. Also the following is disclosed: a pharmaceutical composition and a medicinal product containing the said antibody for the treatment of hemophilia A.

EFFECT: invention makes it possible to effectively treat hemophilia A.

12 cl, 4 dwg, 17 tbl, 3 ex

Description

INCLUDE BY REFERENCE A SEQUENCE LIST

The sequence listing entitled "160163WO02_ST25" is 198 kilobytes and was generated on January 30, 2018 and is incorporated herein by reference.

BACKGROUND OF THE INVENTION

In patients with coagulopathy, such as people with hemophilia A and B, various steps of the coagulation cascade are dysfunctional, for example due to the absence or insufficient amount of functional clotting factor present. This dysfunction of one part of the coagulation cascade leads to insufficient blood clotting and potentially life-threatening bleeding or damage to internal organs such as joints.

Blood clotting factor VIII (FVIII) deficiency, commonly referred to as hemophilia A, is a congenital bleeding disorder affecting approximately 420,000 people worldwide, of whom approximately 105,000 are currently diagnosed.

Patients with hemophilia A may receive clotting factor replacement therapy, such as exogenous FVIII. Traditional treatment consists of replacement therapy given as prophylaxis or on demand to treat bleeding episodes. Until recently, prophylactic treatment for a patient with severe hemophilia A consisted of up to three intravenous injections per week of either plasma-derived FVIII, recombinant FVIII, or long-acting variants thereof.

However, such patients are at risk of developing neutralizing antibodies, called inhibitors, to such exogenous factors, rendering previously effective therapy ineffective. Hemophilia A in patients with inhibitors is a non-limiting example of a coagulopathy that is partly congenital and partly acquired. Patients who develop FVTII inhibitors cannot be treated with conventional replacement therapy. Recently, a new drug, hemlibra, was approved for subcutaneous prophylactic treatment of hemophilia A in the presence of inhibitors. Exogenous blood coagulation factors can only be administered intravenously, which creates significant inconvenience and discomfort for patients. For example, infants and young children may require surgical placement of intravenous catheters into a chest vein to ensure venous access. This puts them at high risk of developing bacterial infections. Thus, even with the advent of Hemlibra, there is a need for an alternative subcutaneous treatment for hemophilia in the presence of inhibitors.

In a bleeding individual, coagulation is initiated by the formation of the tissue factor/factor VIIa (TF/FVIIa) complex when extravascular TF is exposed to activated FVII (FVIIa) in the blood. The formation of the TF/FVIIa complex leads to the activation of factor X (FX) to activated factor Xa (FXa), which, together with activated factor V (FVa), leads to the formation of a limited amount of thrombin, which in turn activates platelets. Activated platelets support the assembly of a tenase complex consisting of activated factor VIII (FVIIIa) and activated factor IX (FIXa). The tenase complex is a very efficient catalyst for FX activation, and the FXa generated in this second step serves as the active protease in the FVa/FXa prothrombinase complex, which is responsible for the final thrombin burst. Thrombin breaks down fibrinogen to form fibrin monomers, which polymerize to form a fibrin network that seals the leaking vessel and stops bleeding. A rapid and extensive thrombin burst is a prerequisite for the formation of a solid and stable fibrin clot.

Inadequate FXa production and decreased thrombin production caused by decreased or absent FVIII activity are the underlying causes of bleeding diathesis in patients with hemophilia A.

As mentioned, the proteolytic conversion of FX to its enzymatically active form FXa can be achieved using a proprietary FX activating complex containing FIXa and its cofactor FVIIIa. Cofactor binding increases the enzymatic activity of FIXa by approximately five orders of magnitude and is believed to occur through multiple mechanisms described by Scheiflinger et al. (2008) J Thromb Haemost, 6:315–322. Notably, FVIIIa was found to stabilize the conformation of FIXa, which has increased proteolytic activity towards FX (Kolkman JA, Mertens K (2000) Biochemistry, 39:7398-7405, Brandstetter H (2009) Biol Chem, 390:391–400). Based on this observation and recognizing that antibodies are versatile binding proteins capable of mimicking a variety of protein-protein interactions, Scheiflinger et al. screened for agonistic antibodies to FIXa, characterized by the ability to enhance FX activation by FIXa in the presence of a phospholipid surface and calcium, but in the absence of the natural cofactor FVIIIa. In a screen of supernatants of 5280 hybridomas, 88 were found to produce antibodies exhibiting varying degrees of FIXa agonist activity, see EP 1220923 B1 and EP 1660536 B1. Regarding the kinetics of FX activation and the ability to stimulate thrombin formation in human plasma with FVIII deficiency, EP 1660536 B1 consistently lists 224F3 as the most effective antibody (see, for example, paragraphs 0060 and 0062).

ACE910 or emicizumab (trade name Hemlibra®) is a humanized bispecific monoclonal antibody to FIX(a)/FX(a) developed by Chugai Pharmaceutical for the treatment of hemophilia A. ACE910 is designed to mimic the function of the FVIII cofactor (see Sampei et al.: ( 2013) PLoS One, 8, e57479 and WO 2012067176).

There are still many unmet medical needs in individuals with hemophilia in particular, and in subjects with types of coagulopathy in general, and the present invention relates to improved compounds capable of replacing FVIII and thus useful for the treatment of coagulopathy such as hemophilia A .

SHORT DESCRIPTION

The present invention relates to compounds that serve as replacements for coagulation factor VIII (FVIII) in patients suffering from coagulopathy, and in particular in patients who lack functional FVIII, such as patients with hemophilia A, including patients with hemophilia A with inhibitors .

Therefore, one aspect of the present invention relates to compounds capable of enhancing the production of FXa and thereby partially or completely restoring coagulation in patients who do not have FVIII.

In one aspect, the compound is an antibody. In one such aspect, the compound is a multispecific antibody, such as a bispecific antibody.

In one particular aspect, the present invention provides procoagulant antibodies that serve as a replacement for FVIII in patients who do not have FVIII, such as patients with hemophilia A.

In one such aspect, the antibody binds to FIXa and increases its enzymatic activity towards FX without necessarily also binding to FX.

In one aspect, the present invention provides a procoagulant antibody that binds FX, including bispecific procoagulant antibodies that increase the enzymatic activity of FIXa toward FX and the extent of FX binding.

In one aspect, the present invention provides a procoagulant bispecific antibody that is capable of binding to the blood clotting factors FIX/FIXa and FX/FXa.

In one aspect, the antibody is human or humanized.

An additional aspect of the present invention relates to individual antibodies or antigen binding fragments thereof that are part of a procoagulant antibody, such as a particular anti-FIX or anti-FIXa antibody or antigen binding fragment thereof. An additional aspect of the present invention relates to individual antibodies or antigen binding fragments thereof that are part of a procoagulant antibody, such as a particular anti-FX or anti-FXa antibody or antigen binding fragment thereof.

An additional aspect of the present invention relates to the manufacture of antibodies and their intermediates disclosed herein.

An additional aspect of the present invention relates to an antibody that competes with a procoagulant antibody or antigen binding fragment thereof disclosed herein for binding to FIX/FIXa.

An additional aspect of the present invention relates to a procoagulant antibody that competes with an antibody or antigen binding fragment thereof disclosed herein for binding to FX/FXa.

Another further aspect of the present invention relates to a pharmaceutical composition containing a procoagulant antibody disclosed herein, formulated to deliver said antibody for the prevention and/or treatment of coagulopathy.

An additional aspect of the present invention is directed to the procoagulant antibodies disclosed herein for the prevention and/or treatment of coagulopathy, a disease accompanying a coagulopathy, or a disease caused by a coagulopathy.

The present invention may also solve additional problems that will be apparent from the disclosure of the illustrative embodiments.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Figure 1 shows the aligned sequences of SEQ ID NO: 3-188, in which the complementarity determining regions 1, 2 and 3 (CDR1, CDR2 and CDR3) of the heavy chain and light chain variable domains were highlighted sequentially in boxes.

Figure 2 shows thrombin generation test (TGT) data for the bispecific antibodies mAb4-7761, mAb4-7762, mAb4-7789, mAb5-0057, mAb5-1409 and ACE910 in tissue factor-activated platelet-poor plasma of human hemophilia A (HA-PPP) ). The experiment was performed as described in Example 17. The dotted and dotted lines indicate the peak level (nM) of thrombin observed in the absence of anti-FVIII antibody in HA-PPP and normal PPP, respectively, and their standard deviation, indicated by the dotted lines. The mAb4-7761, mAb4-7762, mAb4-7789, mAb5-0057 and mAb5-1409 profiles are indicated by upward-pointing triangles, while the ACE910 profile is indicated by downward-pointing triangles. Exp. A - D refer to independent experiments; in each of these experiments, the peak thrombin level at each antibody concentration is the mean ± standard deviation of at least three independent runs.

Figure 3 shows thrombin generation test (TGT) data for the bispecific antibodies mAb5-0057, mAb5-1409 and ACE910 in tissue factor-activated human hemophilia A platelet-rich plasma (HA-PRP). The experiment was performed as described in Example 17. The dotted and dotted lines indicate the peak level (nM) of thrombin observed in the absence of anti-FVIII antibody in HA-PRP and normal PRP, respectively, and their standard deviation, indicated by the dotted lines. The mAb5-0057 and mAb5-1409 profiles are indicated by upward-pointing triangles, while the ACE910 profile is indicated by downward-pointing triangles. Results are presented as mean ± standard deviation from four independent experiments.

Figure 4 shows thrombin generation test (TGT) data for monovalent partial antibodies mAb1-9016 and 224F3 in human platelet-poor plasma activated by factor XIa. The experiment was performed as described in Example 18. The dotted lines indicate the mean peak level (nM) of thrombin observed in the absence of antibody and its standard deviation (±1 SD) indicated by the dotted lines. The profiles of the OA variants mAb1-9016 and 224F3 (mAb1-1582) are indicated by triangles pointing up and down, respectively.

BRIEF DESCRIPTION OF SEQUENCES

SEQ ID NO: 1 is the amino acid sequence of human coagulation factor IX.

SEQ ID NO: 2 is the amino acid sequence of human blood clotting factor X.

SEQ ID NO: 3-188 are the variable heavy chain ( _VH ) and variable light domain ( _VL ) sequences of the anti-FIX and anti-FX monoclonal antibodies (mAbs) described herein. IDs for the corresponding partial (OA) antibodies, as well as some bispecific antibodies, are also given in the table. The CDR1-3 sequences are highlighted by boxes in Figure 1.

Overview of antibody abbreviations, targets, and SEQ ID NOs for the corresponding V _H and V _L sequences.

The first column (“ID for OA or bispecific antibody”) contains abbreviations for monovalent partial (OA) antibodies and/or bispecific antibodies. The second column (“ID mAb”) provides the abbreviations for the corresponding moiety antibodies (for bispecific antibodies, the first antibody mentioned in the second column is the anti-FIX/FIXa antibody and the second is the anti-FX/FXa antibody). The fourth (“SEQ ID NO (V _H )”) and fifth (“SEQ ID NO (V _L )”) columns represent the SEQ ID NO for the sequences V _H and V _L , respectively, with the first SEQ ID NO in each column representing the antibody to FIX/FIXa, and the second is an antibody to FX/FXa.

DESCRIPTION

In subjects with coagulopathy, such as people with hemophilia A, the coagulation cascade becomes dysfunctional due to the absence or insufficient amount of functional FVIII present. This dysfunction of one part of the coagulation cascade leads to insufficient blood clotting and potentially life-threatening bleeding or damage to internal organs such as joints. The present invention relates to compounds that serve as replacements for coagulation factor VIII (FVIII) in patients suffering from coagulopathy, and in particular in patients who lack functional FVIII, such as patients with hemophilia A, including patients with hemophilia A with inhibitors . In one aspect, such a compound is an antibody.

In particular, the inventors of the present invention have unexpectedly identified antibodies that mimic the activity of the FVIII cofactor with high potency and efficiency.

In one particular aspect, the present invention provides procoagulant antibodies that serve as a replacement for FVIII in patients who do not have functional FVIII, such as patients with hemophilia A.

In one such aspect, procoagulant antibodies bind to factor IXa (FIXa) and increase its enzymatic activity towards factor X (FX), without necessarily also binding to FX. In one such aspect, the antibodies of the present invention are bispecific antibodies capable of binding to FIX/FIXa and FX.

Clotting factor IX

FIX is a vitamin K-dependent coagulation factor with structural similarities to factor VII, prothrombin, factor X, and C protein. The circulating form of zymogen consists of 415 amino acids divided into four distinct domains, including an N-terminal γ-carboxyglutamic acid (Gla)-rich domain, two EGF domains, and a C-terminal domain with trypsin-like serine protease properties. FIX circulates in plasma as a single-chain zymogen (SEQ ID NO: 1). Activation of FIX occurs by limited proteolysis at Arg145 and Arg180 to release the activation peptide (residues 146 to 180 of SEQ ID NO: 1). Thus, activated FIX (FIXa) consists of residues 1-145 of SEQ ID NO: 1 (light chain) and residues 181-415 of SEQ ID NO: 1 (heavy chain).

Circulating FIX molecules thus include the FIX zymogen and the activated form of FIX, which are collectively referred to herein as FIX and FIXa with reference to SEQ ID NO: 1.

Activated factor IX is referred to as factor IXa or FIXa. The term "FIX (SEQ ID NO: 1) and/or its activated form (FIXa)" may also be referred to as "FIX/FIXa" or "FIX(a)".

FIXa is a trypsin-like serine protease that plays a key role in hemostasis by generating as part of the tenase complex most of the factor Xa necessary to maintain proper thrombin formation during coagulation.

FIX is represented herein by the sequence at SEQ ID NO: 1, corresponding to the allelic form of human Ala148 FIX (Anson et al. EMBO J. 1984 3:1053-1060; McGraw et al, Proc Natl Acad Sci USA. 1985 82:2847-2851 (Graham et al. Am J Hum Genet 1988 42:573-580). In the present invention, FIX is intended to cover all naturally occurring FIX variants, such as the T148 variant (Uniprot ID P00740).

Clotting factor X

FX is a vitamin K-dependent coagulation factor with structural similarities to factor VII, prothrombin, FIX, and C protein. Human FX zymogen contains four distinct domains, including an N-terminal gamma-carboxyglutamic acid (Gla)-rich domain, two EGF domains, and a C-terminal domain with trypsin-like serine protease properties. FX circulates in the blood plasma as a double-chain zymogen comprising residues 1-139 of SEQ ID NO: 2 (light chain) and residues 143-448 of SEQ ID NO: 2 (heavy chain). Activation of FX occurs through limited proteolysis at Arg194, which leads to the release of the activation peptide (aa 143-194). Thus, activated FX (FXa) consists of residues 1-139 of SEQ ID NO: 2 (light chain) and residues 195-448 of SEQ ID NO: 2 (activated heavy chain). Circulating FX molecules thus include the FX zymogen and the activated form of FX, which are herein referred to as FX and FXa, respectively, with reference to SEQ ID NO: 2. In the present invention, FX is intended to cover all naturally occurring FX variants. The term "FX (SEQ ID NO: 2) and/or its activated form (FXa)" may also be referred to as "FX/FXa" or "FX(a)".

Antibodies

The term “antibody” as used herein refers to a protein derived from an immunoglobulin sequence that is capable of binding to an antigen or part thereof. The term antibody includes, without limitation, full-length antibodies of any class (or isotype), that is, IgA, IgD, IgE, IgG, IgM and/or IgY. The term antibody includes, without limitation, antibodies that are bivalent, such as bispecific antibodies.

Natural full-length antibodies contain at least four polypeptide chains: two heavy chains (HC) and two light chains (LC), which are linked by disulfide bonds. In some cases, natural antibodies contain fewer than four chains, as is the case with IgNARs found in Chondrichthyes. One class of immunoglobulins of particular pharmaceutical interest is IgG. In humans, the IgG class can be divided into four subclasses IgG1, IgG2, IgG3 and IgG4 based on the sequence of their heavy chain constant regions. Light chains can be divided into two types: kappa and lambda chains, based on differences in their sequence compositions. IgG molecules consist of two heavy chains linked by two or more disulfide bonds and two light chains, each attached to the heavy chain by a disulfide bond. An IgG heavy chain may contain a heavy chain variable domain ( _VH ) and up to three heavy chain constant domains ( _CH ): _CH1 , _CH2 and _CH3 . A light chain may contain a light chain variable domain ( _VL ) and light chain constant domain ( _CL ). The V _H and V _L regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) or hypervariable regions (HvRs), interspersed with more conserved regions called framework regions (FRs). The V _H and V _L domains typically consist of three CDRs and four FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable domains of the heavy and light chains, containing hypervariable regions (CDRs), form a structure that is capable of interacting with the antigen, while the constant region of the antibody can mediate binding of the immunoglobulin to host tissues or factors, including, but not limited to, various cells immune system (effector cells), Fc receptors and the first component, C1q, from the C1 complex of the classical complement system.

The antibodies of the present invention may be monoclonal antibodies (mAbs) in the sense that they are a set of unique heavy and light chain variable domain sequences expressed by a single B cell or clonal population of B cells. Antibodies of the present invention can be prepared and purified using various methods known to one skilled in the art. For example, antibodies can be obtained from hybridoma cells. Antibodies can be produced by expanding B cells. Antibodies or fragments thereof can be expressed recombinantly in mammalian or microbial expression systems or by in vitro translation. Antibodies or fragments thereof can also be recombinantly expressed as molecules associated with the cell surface, for example, using phage display, bacterial display, yeast display, mammalian cell display, or ribosomal or mRNA display.

Antibodies of the present invention can be isolated. The term "isolated antibody" refers to an antibody that has been separated from and/or extracted from other component(s) in the environment in which it was produced and/or which has been purified from a mixture of components present in the environment in which it was produced.

Certain antigen-binding antibody fragments may be suitable in the context of the present invention, since it has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. The term "antigen-binding fragment" of an antibody refers to one or more antibody fragments that retain the ability to specifically bind or recognize an antigen, such as FIX/FIXa, FX/FXa, or other target molecule, as described herein. Examples of antigen binding fragments include, but are not limited to, Fab, Fab', Fab ₂ , Fab' ₂ , Fv (typically a combination of the V _L and V _H domains from one arm of the antibody), single chain Fv (scFv); see, for example, Bird et al. Science 1988; 242:423-426 and Huston et al. PNAS 1988; 85:5879-5883), dsFv, Fd (usually V _H and _CH 1 domain), monovalent molecules containing both one V _H and one V _L domain; minibodies, diabodies, tribodies, tetrabodies and kappa bodies (see, for example, Ill et al (1997) Protein Eng 10:949-57); and one or more isolated CDRs or functional paratopes, wherein the isolated CDRs or antigen-binding residues or polypeptides can be linked or linked to each other to form a functional antibody fragment. These antibody fragments can be prepared using conventional techniques known to those skilled in the art, and the fragments can be screened for utility in the same manner as intact antibodies.

"Fab fragments" of an antibody, including "Fab" and "Fab' ₂ " fragments, can be produced from an antibody by heavy chain cleavage at the hinge region at the N-terminal or C-terminal side, respectively, of the hinge cysteine residues connecting the heavy chains of the antibody. The "Fab" fragment comprises a light chain variable and constant domain and a heavy chain variable and _CH 1 domain. Fab' ₂ fragments contain a pair of Fab' fragments that are typically covalently linked by their hinge cysteines. Fab' is typically prepared from the Fab' ₂ fragment by cleavage of the hinge disulfide bonds connecting the heavy chains in Fab' ₂ . Other chemical bonds other than disulfide bonds of antibody fragments are also known in the art. The Fab fragment retains the ability of the parent antibody to bind to its antigen, potentially with lower affinity. Fab' ₂ fragments are capable of bivalent binding, whereas Fab and Fab' fragments can only bind monovalently. As a rule, Fab fragments lack the _CH2 and _CH3 constant domains, i.e. Fc part, where interaction with Fc receptors and C1q can occur. Thus, Fab fragments generally lack effector functions. Fab fragments can be prepared by methods known in the art or by enzymatic digestion of the antibody, for example using papain to produce Fab or pepsin to produce Fab' ₂ , or Fab fragments including Fab, Fab', Fab' ₂ , can be produced recombinantly using techniques that are well known to one skilled in the art.

An "Fv" fragment (variable fragment) is an antibody fragment that contains a complete antigen recognition and binding site and typically contains one heavy chain variable domain and one light chain variable domain in an association that may be covalent in nature, such as a single chain variable domain fragment (scFv). It is in this configuration that the three hypervariable regions of each variable domain interact to form an antigen binding site on the surface of the V _H -V _L dimer. Collectively, the six hypervariable regions or a subset thereof provide the antibody with antigen binding specificity.

A "single chain Fv" or "scFv" antibody contains the V _H and V _L domains of the antibody, where these domains are present on the same polypeptide chain. Typically, the Fv polypeptide additionally contains a polypeptide linker between the V _H and V _L domains, which allows the scFv to form the necessary structure for antigen binding. For a review regarding scFv, see Pluckthun, 1994, in: The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315.

A "single chain Fab" or "scFab" antibody contains the V _H , _CH 1, V _L and C _L domains of the antibody, where these domains are present on a single polypeptide chain. Typically, the Fab polypeptide additionally contains a polypeptide linker between the V _H and C _L or V _L and C _H 1 domains, which allows the scFab to form the necessary structure for antigen binding (Koerber et al. (2015) J Mol Biol. 427:576-86 ).

The term "diabodies" refers to small antibody fragments with two antigen-binding regions, the fragments containing a heavy chain variable domain ( _VH ) joined to a light chain variable domain ( _VL ) in the same polypeptide chain ( _VH and _VL ) . By using a linker that is too short to allow pairing between two variable domains on the same chain, the variable domains are forced to pair with complementary domains on the other chain, creating two antigen-binding sites.

The expression "linear antibodies" refers to the antibodies described in Zapata et al. (1995) Protein Eng. 8:1057–1062. Briefly, these antibodies contain a pair of tandem Fd segments (V _H -C _H 1-V _H -C _H 1), which together with complementary light chain polypeptides form a pair of antigen binding regions. Linear antibodies can be bispecific or monospecific.

Antibody fragments can be produced using conventional recombinant or protein engineering techniques, and the fragments can be screened for binding to FIX and its activated form, FX or other functional element in the same manner as intact antibodies.

Antibody fragments of the present invention can be produced by truncation, for example by removing one or more amino acids from the N- and/or C-terminus of the polypeptide. Fragments can also be generated by one or more internal deletions.

An antibody of the present invention may be or may contain an antibody fragment or variant of any of the antibodies disclosed herein. An antibody of the present invention may be or may contain an antigen-binding portion of one of these antibodies or variants thereof. For example, an antibody of the present invention may be a Fab fragment of one of these antibodies or variants thereof, or it may be a single chain antibody derived from one of these antibodies or a variant thereof. Also, the antibody of the present invention may be a combination of a full-length antibody and a fragment thereof.

As used herein, the term “partial” refers to a monovalent antibody of a particular type, consisting of an antibody heavy chain, a truncated heavy chain lacking the Fab region, and one light chain.

As used herein, the term “monospecific” antibody refers to an antibody that is capable of binding to one specific epitope (including, without limitation, bivalent antibodies).

As used herein, the term “bispecific” antibody refers to an antibody that is capable of binding to two different antigens or two different epitopes on the same antigen.

As used herein, the term “trispecific” antibody refers to an antibody that is capable of binding to three different antigens, or three different epitopes on the same antigen, or three different epitopes present on two different antigens.

As used herein, the term “multispecific” antibody refers to an antibody that is capable of binding to two or more different antigens or two or more different epitopes on the same antigen. Thus, multispecific antibodies include bi- and trispecific antibodies.

Bispecific antibodies in full-length IgG format can be produced by fusing two separate hybridomas to form a hybrid quadromy, which produces an antibody mixture including a fraction of bispecific heterodimerizing antibodies (Chelius D. et al.; MAbs. 2010 May-Jun; 2(3):309 -319). Bispecific heterodimerizing antibodies can alternatively be produced using recombinant techniques. Heterodimerization can also be achieved by designing a dimerization interface in the Fc region to promote heterodimerization. One example of this is the so-called "overhang-to-hole" mutations, where sterically bulky side chains (overhangs) are introduced into one Fc that matches sterically small side chains (holes) on the opposite Fc, thereby creating steric complementarity that promotes heterodimerization . Other ways to design interfaces on the Fc for heterodimerization are electrostatic complementarity, fusion with non-IgG heterodimerization domains, or use of the natural phenomenon of human IgG4 Fab arm exchange to control heterodimerization. Examples of heterodimerized bispecific antibodies are well described in the literature, for example (Klein C, et al.; MAbs. 2012 Nov-Dec; 4(6):653-663). Particular attention should be paid to the light chains in heterodimeric antibodies. Correct pairing of LC and HC can be achieved by using a common light chain. Again, engineering of the LC/HC interface can be used to promote heterodimerization or engineering of light chain crossover, as in CrossMabs. Reassembly of antibodies in vitro under mild reducing conditions from two separate IgGs containing the appropriate mutations can also be used to produce bispecific antibodies (e.g., Labrijn et al., PNAS, 110, 5145-5150 (2013)). It is also reported that nature's Fab arm exchange mode ensures proper light chain pairing.

Multispecific antibody molecules can also be expressed recombinantly as fusion proteins combining natural IgG modules to form multispecific and multivalent antibody derivatives, as described in the literature. Examples of fusion antibodies are DVD-Ig, IgG-scFV, diabodies, DART, etc. Specific tags for detection or purification, fragments to extend half-life, or other components can be introduced into the fusion proteins. Additional non-IgG attributes can also be introduced into the fusion proteins. Full-length bispecific antibodies based on Fc heterodimerization are generally referred to as asymmetric IgG, regardless of LC pairing methodology.

In general, bispecific antibodies can be produced in various molecular formats, as described by Brinkmann et al. (Brinkmann et al. The making of bispecific antibodies. Mabs 9, 182-212 (2017)).

Multispecific antibody molecules can also be prepared by chemically conjugating or conjugating individual full-length IgGs or conjugating IgG fragments to form multispecific and multivalent antibody derivatives as described in the literature. Examples are chemically coupled Fab fragments, IgG dimer, etc. Conjugated proteins can be loaded with specific tags for detection or purification, molecules to extend half-life, or other components. Additional non-IgG polypeptides can also be incorporated into the fusion proteins. Polyspecific molecules can also be produced by combining recombinant and chemical methods, including those described above.

In one aspect, the antibody of the present invention is a chimeric antibody, a human antibody, or a humanized antibody. Such an antibody can be produced using, for example, suitable antibody display or immunization platforms or other suitable platforms or methods known in the art. As used herein, the term “human antibody” is intended to refer to antibodies having variable domains in which at least a portion of the framework region and/or at least a portion of the CDR region are derived from human germline immunoglobulin sequences. For example, a human antibody may have variable domains in which both framework and CDR regions are derived from human germline immunoglobulin sequences. In addition, if the antibody contains a constant region, then the constant region or part thereof is also derived from human germline immunoglobulin sequences. The human antibodies of the present invention may include amino acid residues not encoded by human germline immunoglobulin sequences (eg, mutations introduced by random or site-specific mutagenesis in vitro or somatic mutation in vivo).

Such a human antibody may be a human monoclonal antibody. Such a human monoclonal antibody may be produced by a hybridoma that includes a B cell derived from a transgenic non-human animal, such as a transgenic mouse, having a genome comprising human immunoglobulin heavy and light chain gene segment repertoires fused to an immortalized cell.

Human antibodies can be isolated from sequence libraries constructed from samples of human germline sequences, further modified by introducing natural and synthetic sequence diversity.

Human antibodies can be obtained by immunizing human lymphocytes in vitro followed by transformation of the lymphocytes with the Epstein-Barr virus.

Human antibodies can be produced by recombinant methods known in the art.

The term “human antibody derivative” refers to any modified form of a human antibody, such as a conjugate of an antibody and another agent or antibody.

As used herein, the term “humanized antibody” refers to a human/non-human antibody that contains a sequence (CDR regions or portions thereof) derived from a non-human immunoglobulin. Thus, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from at least the hypervariable region of the recipient are replaced by residues from the hypervariable region of an antibody from a non-human species (donor antibody), such as a mouse, rat, rabbit, or primate, non-human that has the required sequence specificity, affinity, composition and functionality. In some cases, framework (FR) residues of human immunoglobulin are replaced with corresponding non-human residues. An example of such a modification is the introduction of one or more so-called reverse mutations, which are usually amino acid residues derived from the donor antibody. Humanization of an antibody can be accomplished using recombinant techniques known to one skilled in the art (see, for example, Antibody Engineering, Methods in Molecular Biology, vol. 248, edited by Benny K. Lo). Suitable human recipient frameworks for both light and heavy chain variable domains can be identified, for example, by sequence or structure homology. Alternatively, established recipient scaffolds can be used, for example based on knowledge of the structure, biophysical and biochemical properties. Recipient scaffolds can be derived from the germ line or derived from the mature antibody sequence. CDR regions from a donor antibody can be transferred using CDR grafting. The CDR-grafted humanized antibody can be further optimized, for example, with respect to affinity, functionality and biophysical properties, by identifying essential framework positions where reintroduction (back mutation) of an amino acid residue from the donor antibody has a beneficial effect on the properties of the humanized antibody. In addition to back mutations derived from donor antibodies, a humanized antibody can be constructed by introducing germline residues into CDRs or framework regions, removal of immunogenic epitopes, site-directed mutagenesis, affinity maturation, etc.

Moreover, humanized antibodies may contain residues that are not found in either the recipient antibody or the donor antibody. These modifications are made to further improve the characteristics of the antibody. In general, a humanized antibody will contain at least one, typically two, variable domains in which all or substantially all of the CDR regions correspond to regions of a non-human immunoglobulin, and in which all or substantially all of the FR residues are residues from human immunoglobulin sequences. The humanized antibody may optionally also contain at least a portion of an immunoglobulin constant region (Fc), typically human immunoglobulin.

The term “humanized antibody derivative” refers to any modified form of a humanized antibody, such as an antibody-chemical conjugate or an antibody-another antibody conjugate.

As used herein, the term “chimeric antibody” refers to an antibody containing antibody portions derived from two or more species. For example, genes encoding such an antibody include genes encoding variable domains and genes encoding constant domains originating from two different species. For example, genes encoding the variable domains of a murine monoclonal antibody can be linked to genes encoding the constant domains of a human-derived antibody.

The crystallizing fragment ("Fc region"/"Fc domain") of an antibody is the C-terminal region of the antibody, which contains the hinge and constant domains C _H 2 and C _H 3. The Fc domain can interact with cell surface receptors called Fc -receptors, as well as with some proteins of the complement system. The Fc region allows antibodies to interact with the immune system. In one aspect of the present invention, antibodies can be engineered to include modifications to the Fc region, typically to alter one or more of its functional properties, such as serum half-life, complement fixation, Fc receptor binding, protein stability and/or antigen- dependent cellular cytotoxicity or lack thereof, among others. In addition, the antibody of the present invention can be chemically modified (eg, one or more chemical moieties can be attached to the antibody) or modified to alter its glycosylation, again to alter one or more functional properties of the antibody. An IgG1 antibody may carry a modified Fc domain containing one or more and possibly all of the following mutations that will result in decreased affinity for certain Fc gamma receptors (L234A, L235E, and G237A) and decreased C1q-mediated complement fixation (A330S and P331S), respectively (numbering of residues in accordance with the EU index). Alternatively, other amino acid substitutions and combinations thereof and combinations thereof known in the art may be used to produce altered (decreased or increased) Fc gamma receptor binding.

The antibody isotype of the present invention may be IgG, such as IgG1, such as IgG2, such as IgG4. If desired, the antibody class can be “switched” using known techniques. For example, an antibody that was originally produced as an IgM molecule can be class switched to an IgG antibody. Class switching techniques can also be used to convert one IgG subclass to another, for example: from IgG1 to IgG2 or IgG4; from IgG2 to IgG1 or IgG4; or from IgG4 to IgG1 or IgG2. It is also possible to design antibodies to produce constant region chimeric molecules by combining regions from different IgG subclasses.

In one embodiment, the hinge region of the antibody is modified such that the number of cysteine residues in the hinge region is changed, such as increased or decreased. This approach is further described, for example, in US patent No. 5677425 from Bodmer et al.

The constant region can be modified to stabilize the antibody, for example to reduce the risk of the bivalent antibody separating into semi-antibodies. For example, in the lgG4 constant region, residue S228 (according to the EU numbering index and Kabat S241) can be mutated to a proline residue (P) to stabilize the formation of a disulfide bridge between the heavy chains in the hinge (see, for example, Angal et al. Mol Immunol. 1993 ; 30:105-8).

Antibodies or fragments thereof can be defined in terms of their complementarity determining regions (CDRs). The term “complementarity determining region” or “hypervariable region” as used herein refers to regions of an antibody that contain amino acid residues involved in antigen binding. The hypervariability region or CDR can be identified as the regions with the highest variability in the amino acid alignment of antibody variable domains. Databases can be used to identify CDRs, such as the Kabat database, where CDRs are, for example, defined as containing amino acid residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) of the light chain variable domain and 31 -35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; (Kabat et al. 1991; Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). Alternatively, CDRs can be defined as those residues from the "hypervariable loop" (residues 26-33 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1) 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk, J. Mol. Biol. 1987;196:901-917). Typically, numbering of amino acid residues in this region is performed in the manner described in Kabat et al. higher. Phrases such as “Kabat position,” “Kabat residue,” and “Kabat residue” as used herein refer to this numbering system for heavy chain variable domains or light chain variable domains. When using the Kabat numbering system, the actual linear amino acid sequence of the peptide may contain fewer or additional amino acids corresponding to the truncation or insertion into the framework (FR) or CDR of the variable domain. For example, the heavy chain variable domain may include amino acid insertions (residues 52a, 52b and 52c according to Kabat) after residue 52 of the H2 CDR and inserted residues (e.g. residues 82a, 82b and 82c, etc. according to Kabat) after residue 82 of the heavy chain FR. Kabat residue numbering can be determined for a given antibody by aligning regions of homology between the antibody sequence and the Kabat numbered “standard” sequence.

The term “framework region” or “FR” residues refers to those V _H or V _L amino acid residues that are not in the CDR as defined herein.

An antibody of the present invention may comprise a CDR region from one or more specific antibodies disclosed herein.

The term "procoagulant antibody" refers to an antibody that enhances blood clotting, for example, by accelerating the blood clotting process and/or increasing the enzymatic activity of one or more clotting factors.

The term “procoagulant activity” refers to the ability of a compound, such as an antibody, to enhance blood clotting, for example, by accelerating the blood clotting process and/or increasing the enzymatic activity of one or more clotting factors.

The term "antigen" (Ag) refers to a molecular entity used to immunize an immunocompetent vertebrate animal to produce an antibody (Ab) that recognizes the Ag. As used herein, Ag is defined more broadly and is generally intended to include target molecules that are specifically recognized by an Ab, thus including fragments or mimics of a molecule used in an immunization process or other process, such as phage display, used to produce an Ab.

The term "epitope" as used herein is defined in the context of the molecular interaction between an "antigen-binding polypeptide", such as an antibody (Ab), and its corresponding antigen (Ag). Typically, "epitope" refers to the region or region on Ag to which Ab binds, i.e. to the area or site of physical contact with Ab. Physical contact can be determined using various criteria (e.g. cut-off distance 2-6 , such as 3 , such as 3.5 , such as 4 , such as 4.5 , such as 5 , or solvent accessibility) for atoms in Ab and Ag molecules.

FIX/FIXa and FX/FXa may contain a number of different epitopes, which may include, but are not limited to, (1) linear peptide epitopes, (2) conformational epitopes, which consist of one or more non-contiguous amino acids located adjacent to each other in the mature conformation FIX/FIXa or FX/FXa; and (3) epitopes, which consist, in whole or in part, of molecular structures covalently attached to FIX/FIXa or FX/FXa, such as carbohydrate groups.

The epitope for a given antibody (Ab)/antigen (Ag) pair can be described and characterized at different levels of detail using various experimental and computational epitope mapping techniques. Experimental methods include mutagenesis, x-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, hydrogen-deuterium exchange mass spectrometry (HDX-MS) and various competitive binding methods; methods that are known from the prior art. Because each method is based on a unique principle, the description of an epitope is closely related to the method by which it was determined. Thus, depending on the epitope mapping method used, the epitope for a given Ab/Ag pair can be described in different ways.

In the context of an X-ray crystal structure defined by the spatial coordinates of the complex between an Ab, such as a Fab fragment, and its Ag, the term epitope herein, unless otherwise noted or inconsistent with context, is specifically defined as FIX/FIXa or FX residues , characterized by the presence of a heavy atom (i.e. an atom other than hydrogen) at a distance of 3.5 from a heavy atom in Ab.

Epitopes described at the amino acid level, such as those determined by X-ray structure, are considered identical if they contain the same set of amino acid residues. Epitopes are considered to overlap if at least one amino acid residue is common to the epitopes. Epitopes are considered to be distinct (unique) if there are no amino acid residues that are common to the epitopes.

The definition of the term "paratope" is derived from the above definition of "epitope" by changing the perspective. Thus, the term "paratope" refers to the region or site on the Ab to which Ag binds, i.e. through which it comes into physical contact with Ag.

In the context of an X-ray crystal structure defined by the spatial coordinates of a complex between an Ab, such as a Fab moiety, and its Ag, the term paratope as used herein, unless otherwise noted or inconsistent with context, is specifically defined as Ab residues characterized by the presence of heavy atom (i.e. an atom other than hydrogen) at a distance of 3.5 from the heavy atom in FIX/FIXa or FX.

The epitope and paratope for a given antibody (Ab)/antigen (Ag) pair can be identified by conventional methods. For example, the general location of an epitope can be determined by assessing the ability of an antibody to bind to various fragments or variants of FIX/FIXa or FX. The specific amino acids in FIX/FIXa or FX that come into contact with the antibody (epitope) and the specific amino acids in the antibody that come into contact with FIX/FIXa or FX (paratope) can also be determined by conventional methods. For example, the antibody and the target molecule can be combined and the Ab:Ag complex can be crystallized. The crystal structure of the complex can be determined and used to identify specific sites of interaction between the antibody and its target.

Epitopes on an antigen may contain one or more hot spot residues, i.e. residues that are particularly important for interaction with the cognate antibody, and where interactions mediated by the side chain of the specified hot spot residue contribute significantly to the binding energy for the antibody/antigen interaction (Peng et al. (2014) PNAS 111, E2656-E2665 ). Hot spot residues can be identified by testing antigen variants (herein FIX/FIXa and FX) in which individual epitope residues have been replaced, for example by alanine, for binding to the cognate antibody. If replacement of an epitope residue with an alanine has a strong effect on antibody binding, then the epitope residue is considered a hot spot residue and is therefore of particular importance for antibody binding to the antigen.

Antibodies that bind to the same antigen can be characterized in terms of their ability to bind to their common antigen at the same time and can be subjected to "competitive binding"/"categorization". In the context of the present invention, the term "categorization" refers to a method of grouping antibodies that bind to the same antigen. "Categorization" of antibodies can be based on the competitive binding of two antibodies to their common antigen in assays based on standard techniques.

The “category” of an antibody is determined using a reference antibody. If the second antibody is unable to bind to the antigen at the same time as the reference antibody, then the second antibody is said to be in the same "category" as the reference antibody. In this case, the reference and the second antibody competitively bind the same portion of the antigen and are “competing antibodies.” If the second antibody is able to bind to an antigen at the same time as the reference antibody, then the second antibody is said to be in a separate “category.” In this case, the reference and second antibody do not competitively bind the same portion of the antigen and are “non-competing antibodies.”

“Categorization” of antibodies does not provide direct information about the epitope.

Competing antibodies, i.e. antibodies belonging to the same "category" may have identical epitopes, overlapping epitopes, or even separate epitopes. The latter occurs if the reference antibody, bound to its epitope on the antigen, occupies the space necessary for the second antibody to come into contact with its epitope on the antigen (“steric hindrance”). Non-competing antibodies usually have distinct epitopes. Thus, in some embodiments, the antibodies of the present invention will bind to the same epitope as at least one of the antibodies specifically disclosed herein.

Competition assays to determine whether an antibody competes for binding with the anti-FIX/FIXa antibody or anti-X antibody disclosed herein are known in the art. Exemplary competition assays include immunoassays (eg, ELISA assays, RIA assays), surface plasmon resonance assays (eg, using a BIAcore™ instrument), biolayer interferometry (ForteBio®), and flow pitometry.

Typically, a competition assay involves the use of a solid surface bound or cell surface expressed antigen, a test FIX or FIXa binding antibody, and a reference antibody. The reference antibody is labeled, but the test antibody is not labeled. Competitive inhibition is measured by determining the amount of labeled reference antibody bound to a solid surface or cells in the presence of the test antibody. Typically, the antibody being tested is present in excess (eg, 1, 5, 10, 20, 100, 1000, 10,000, or 100,000 times). Antibodies identified as competing in a competition assay (i.e., competing antibodies) include antibodies that bind to the same epitope or overlapping epitopes as the reference antibody and antibodies that bind to an adjacent epitope sufficiently proximal to the epitope that bound to the reference antibody to cause steric interference.

In an exemplary competition assay, a reference anti-FIX or anti-FIXa antibody is biotinylated using commercially available reagents. The biotinylated reference antibody is mixed with serial dilutions of the test antibody or unlabeled reference antibody (self-competition control), resulting in a mixture of different molar ratios (e.g., 1, 5, 10, 20, 100, 1000, 10,000, or 100,000 times) of the test antibody (or unlabeled reference antibody) with labeled reference antibody. The antibody mixture is added to an ELISA plate coated with FIX or FIXa polypeptide. The plate is then washed and horseradish peroxidase (HRP)-streptavidin is added to the plate as a detection reagent. The amount of labeled reference antibody bound to the target antigen is determined after the addition of a chromogenic substrate (for example, TMB (3,3',5,5'-tetramethylbenzidine) or ABTS (2,2''-azino-di-(3-ethylbenzthiazolin- 6-sulfonate)), which are known in the art. Optical density (OD units) readings are obtained using a spectrometer (e.g., SpectraMax® M2 spectrometer (Molecular Devices)). The response (OD units) corresponding to zero percent inhibition is determined from wells without any competing antibody. The response (OD units) corresponding to 100% inhibition, i.e., assay background, is determined from wells without any labeled reference antibody or test antibody. Percentage of inhibition of labeled reference antibody to FIX or FIXa by test antibody (or unlabeled reference antibody) at each concentration is calculated as follows: % inhibition = (1 - (OD units - 100% inhibition) / (0% inhibition - 100% inhibition)) * 100.

One skilled in the art will appreciate that similar assays can be performed to determine whether two or more anti-FX/FXa antibodies share a common binding region, category, and/or competitively bind an antigen. Those skilled in the art will also recognize that competitive analysis can be performed using various detection systems known in the art.

A test antibody competes with a reference antibody for antigen binding if an excess of one antibody (e.g., 1, 5, 10, 20, 100, 1000, 10,000, or 100,000 times) inhibits binding of the other antibody, e.g., by at least 50%, 75 %, 90%, 95% or 99%, as measured in a competitive binding assay.

Unless otherwise stated, competition was determined using a competition ELISA assay as described above.

The term "binding affinity" is used herein as a measure of the strength of a non-covalent interaction between two molecules, such as an antibody or fragment thereof and an antigen. The term binding affinity is used to describe monovalent interactions.

The binding affinity between two molecules, such as an antibody or fragment thereof, and an antigen, via a monovalent interaction, can be quantified by determining the equilibrium dissociation constant (K _D ). K _D can be determined by measuring the kinetics of complex formation and dissociation, for example by the surface plasmon resonance (SPR) method or the isothermal titration calorimetry (ITC) method. The rate constants corresponding to the association and dissociation of a monovalent complex are referred to as the association rate constant k _a (or k _on ) and the dissociation rate constant k _d (or k _off ), respectively. K _D is related to k _a and k _d through the equation K _D =k _d / k _a .

Following the above definition, the binding affinities associated with various molecular interactions, such as comparing the binding affinities of different antibodies for a given antigen, can be compared by comparing K _D values for individual antibody/antigen complexes.

The value of the dissociation constant can be determined directly by well known methods. Standard assays for assessing the binding ability of ligands, such as antibodies, to targets are known in the art and include, for example, ELISA, Western blots, RIA and flow cytometry analysis. The binding kinetics and binding affinity of an antibody can also be assessed using standard assays known in the art, such as SPR. However, it is preferable that isothermal titration calorimetry (ITC) can be used to measure the affinity of the antibody/target interaction, as well as to obtain thermodynamic parameters of the interaction.

A competitive binding assay can be performed in which the binding of an antibody to a target is compared with the binding of the target by another ligand of that target, such as another antibody.

An antibody of the present invention may have a K _D for its target of 1×10 ^-4 M or less, 1×10 ^-5 M or less, 1×10 ^-6 M or less, 1×10 ^-7 M or less, 1×10 ^-8 M or less, or 1×10 ^-9 M or less, or 1×10 ^-10 M or less, 1×10 ^-11 M or less, 1×10 ^-12 M or less, 1×10 ^-13 M or less or 1×10 ^-14 M or less.

The K _D of the antibody of the present invention may be less than 100 μM, such as less than 10 μM, such as less than 1 μM, such as less than 0.9 μM, such as less than 0. 8 µM, such as less than 0.7 µM, such as less than 0.6 µM, such as less than 0.5 µM, such as less than 0.4 µM, such as , less than 0.3 μM, such as less than 0.2 μM, such as less than 0.1 μM.

In one such embodiment, the antibody is a bispecific antibody comprising an anti-FX arm with a KD for FX of less than 100 μM, such as less than 10 μM, such as less than 1 μM, such as less than 0 .9 µM, such as less than 0.8 µM, such as less than 0.7 µM, such as less than 0.6 µM, such as less than 0.5 µM, such as, for example, less than 0.4 μM, such as less than 0.3 μM, such as less than 0.2 μM, such as less than 0.1 μM, such as less than 0, 09 µM, such as less than 0.08 µM, such as less than 0.07 µM, such as less than 0.06 µM, such as less than 0.05 µM, such as , less than 0.04 μM, such as less than 0.03 μM, such as less than 0.02 μM, such as less than 0.01 μM, such as less than 9 nM, such as less than 8 nM, such as less than 7 nM, such as less than 6 nM, such as less than 5 nM, such as less than 4 nM, such as less less than 3 nM, such as less than 2 nM, such as less than 1 nM, such as less than 0.5 nM.

The antibodies and antibody fragments described herein can be combined with other antibodies and antibody fragments known in the art to form bispecific, trispecific, or multispecific antibody molecules. Compounds that mimic FVIII cofactor function have previously been created using other FIX/IXa and FX/Xa binding domains, each of which could potentially replace the FIX/IXa and/or FX/Xa binding domains described herein. Thus, it is clear that the FIX/IXa and FX/Xa binding domains of the present invention are of particular interest as individual molecules, and also as “intermediates” as part of a bi-, tri- or polyspecific antibody containing at least one FIX/IXa and/or FX/Xa binding domain.

The activity of procoagulant antibodies, including bi-, tri- and polyspecific antibodies, can be determined by methods known in the art. Standard tests include thrombin generation test (TGT) in whole blood, clotting time measurements using thromboelastography (TEG), and FXa generation assays.

Identity

The term “identity”, as is known in the art, refers to the relationship between the sequences of two or more polypeptides, determined by comparison of the sequences. In the art, “identity” also means the degree of sequence relatedness between polypeptides, determined by the number of matches between chains of two or more amino acid residues. "Identity" is measured as the percentage of identical matches between the smaller of two or more sequences with gap alignments (if any) using a specific mathematical model or computer program (i.e., "algorithms"). The identity of related polypeptides can be easily calculated by known methods. Such methods include, without limitation, those described in Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A.M., and Griffin, H.G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York, 1991 and Carillo et al. SIAM J. Applied Math. 1988; 48:1073.

Preferred methods for determining identity are designed to provide the greatest match between the tested sequences. Methods for determining identity are described in publicly available computer programs. Preferred methods using computer programs to determine the identity between two sequences include the GCG software package, including GAP (Devereux et al. Nucl. Acid. Res. 1984; 12:387); Genetics Computer Group, University of Wisconsin, Madison, WI), BLASTP, BLASTN, and FASTA (Altschul et al. J. Mol. Biol. 1990; 215:403-410). The BLASTX program is publicly available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul et al. NCB/NLM/NIH Bethesda, MD 20894; Altschul et al. supra). The well-known Smith-Waterman algorithm can also be used to determine identity.

For example, using the GAP computer algorithm (Genetics Computer Group, University of Wisconsin, Madison, WI), two polypeptides for which percent sequence identity is determined are aligned for an optimal match of their respective amino acids (the "match length" as determined by the algorithm). Penalty for introducing a gap (which is calculated as 3 times the average diagonal; "average diagonal" is the average of the diagonal of the comparison matrix used; "diagonal" is the score or number assigned to each perfect amino acid match by a particular comparison matrix) and penalty for continuing the gap (which is usually equal to {fraction (1/10)} of the gap penalty), and a comparison matrix such as PAM 250 or BLOSUM 62 is used in conjunction with the algorithm. Standard comparison matrix (see Dayhoff et al. 1978; Atlas of Protein Sequence and Structure, vol. 5, supp. 3 for the PAM 250 comparison matrix; Henikoff et al. PNAS 1992; 89:10915-10919 for the BLOSUM comparison matrix 62) are also used in the algorithm.

Preferred parameters for comparing peptide sequences include the following: Algorithm: Needleman et al. J. Mol. Biol. 1970; 48:443-453; comparison matrix: BLOSUM 62 from Henikoff et al. PNAS 1992; 89:10915-10919; penalty for introducing a gap: 12, penalty for continuing the gap: 4, similarity threshold: 0.

The GAP program is useful with the above parameters. The above parameters are the default parameters for comparing peptides (along with no gap closure penalty) using the GAP algorithm.

The term "similarity" is a related concept, but unlike "identity" it refers to sequence relationships that include both identical matches and conserved substitution matches. If two polypeptide sequences have, for example, (fraction (10/20)) identical amino acids, and all the rest are non-conservative substitutions, then the percentage of identity and similarity will be 50%. If in the same example there are 5 more positions with conservative substitutions, then the percentage of identity is 25%, and the percentage of similarity will be 75% ((proportion (15/20))). Therefore, in cases where there are conservative substitutions, the degree of similarity between two polypeptides will be higher than the percentage of identity between the two polypeptides.

Pharmaceutical compounds

In another aspect, the present invention relates to compositions and formulations containing the compounds of the present invention, such as the antibodies described herein. For example, the present invention provides a pharmaceutical composition that contains one or more antibodies of the present invention formulated together with a pharmaceutically acceptable carrier.

Accordingly, one object of the present invention is to provide a pharmaceutical composition containing such an antibody which is present in a concentration of from 0.25 mg/ml to 250 mg/ml, and wherein said composition has a pH of from 2.0 to 10.0 . The composition may further contain one or more of a buffer system, a preservative, a tonicity control agent, a chelating agent, a stabilizer or a surfactant, as well as various combinations thereof. The use of preservatives, isotonic agents, chelating agents, stabilizers and surfactants in pharmaceutical compositions is well known to those skilled in the art. Reference may be made to Remington: The Science and Practice of Pharmacy, ^19th edition, 1995.

In one embodiment, the pharmaceutical composition is an aqueous composition. Such a composition is typically a solution or suspension, but may also include colloids, dispersions, emulsions and multiphase materials. The term "aqueous composition" is defined as a composition containing at least 50% w/w water. Likewise, the term "aqueous solution" is defined as a solution containing at least 50% w/w water, and the term "aqueous suspension" is defined as a suspension containing at least 50% w/w water.

In another embodiment, the pharmaceutical formulation is a lyophilized formulation to which the physician or patient adds solvents and/or diluents prior to use.

In a further aspect, the pharmaceutical composition contains an aqueous solution of such antibody and a buffer, wherein the antibody is present at a concentration of 1 mg/ml or higher, and wherein the composition has a pH of from about 2.0 to about 10.0.

Introduction

The compound of the present invention, such as an antibody, can be administered parenterally, for example intravenously, for example intramuscularly, for example subcutaneously. Alternatively, the antibody of the present invention can be administered by a non-parenteral route, for example orally or topically. The antibody of the present invention can be administered prophylactically. The antibody of the present invention can be administered therapeutically (on demand).

Doses

The dose of the compounds to be delivered may be from about 0.01 mg to 500 mg of compound per day, preferably from about 0.1 mg to 250 mg per day, and more preferably from about 0.5 mg to about 250 mg per day, per week, every two weeks or a month as loading and maintenance doses, depending on the severity of the condition. An appropriate dose can also be adjusted for a particular compound based on the properties of that compound, including its in vivo half-life or mean retention time and its biological activity. For example, the compounds to be delivered can be administered in one embodiment once a week, or in another embodiment once every two weeks, or in another embodiment once a month and in any of these embodiments at a dose of, for example, 0. 25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 mg per kg body weight.

Compositions containing the compounds disclosed herein can be administered for prophylactic and/or in some embodiments therapeutic treatment. In therapeutic applications, the compositions are administered to a subject already suffering from a disease, such as any bleeding disorder as described above, in an amount sufficient to cure, alleviate or partially delay the disease and its complications. An amount sufficient to achieve this is defined as a "therapeutically effective amount". As will be appreciated by one skilled in the art, amounts effective for this purpose will depend on the severity of the disease or injury, as well as the weight and general condition of the subject.

Embodiments

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue H256 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue H257 of FIX (SEQ ID NO: 1) or its activated form FIXa.

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue N258 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue K293 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue K301 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue D332 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue R333 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue A334 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue T335 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue L337 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue R338 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue S339 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing the T340 residue of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue K341 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue T343 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue N346 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing the R403 residue of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue Y404 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue N406 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue W407 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing the E410 residue of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope containing residue K411 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope comprising residues L337, R338, S339, T340, K341 and T343 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope comprising residues K301, D332, R333, A334, T335, R338 and N346 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding an epitope comprising residues H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411 of FIX (SEQ ID NO: 1) or its activated form (FIXa).

In one embodiment, the antibody of the present invention is capable of binding FIX (SEQ ID NO: 1) or its activated form (FIXa), where the antibody competes with Fab7236 for binding to FIX.

In one embodiment, the antibody of the present invention is capable of binding FIX (SEQ ID NO: 1) or its activated form (FIXa), where the antibody competes with Fab7237 for binding to FIX.

In one embodiment, the antibody of the present invention is capable of binding FIX (SEQ ID NO: 1) or its activated form (FIXa), where the antibody competes with Fab7238 for binding to FIX.

In one embodiment, the antibody of the present invention binds FIX (SEQ ID NO: 1) or its activated form (FIXa), where the antibody belongs to the same “category” as Fab7236.

In one embodiment, the antibody of the present invention binds FIX (SEQ ID NO: 1) or its activated form (FIXa), where the antibody belongs to the same “category” as Fab7237.

In one embodiment, the antibody of the present invention binds FIX (SEQ ID NO: 1) or its activated form (FIXa), where the antibody belongs to the same “category” as Fab7238.

In one embodiment, an antibody of the present invention is capable of binding FX (SEQ ID NO: 2) or its activated form FXa, where the antibody competes with an antibody containing the CDR of mAb1-6723.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody contains the variable domains from mAb1-6723 in accordance with SEQ ID NO: 21 and SEQ ID NO: 22.

In one embodiment, an antibody of the present invention is capable of specifically binding FX/FXa, wherein the antibody comprises a CDR from mAb1-6723 according to SEQ ID NO: 21 and SEQ ID NO: 22.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as the Fab containing the variable domains from mAb1-6723 in accordance with SEQ ID NO: 21 and SEQ ID NO: 22.

In one embodiment, the antibody of the present invention is capable of specifically binding FX/FXa, where the antibody belongs to the same "category" as mAb1-6723 according to SEQ ID NO: 21 and SEQ ID NO: 22.

In one embodiment, the antibody of the present invention is capable of specifically binding FX/FXa, where the antibody belongs to the same "category" as the antibody or Fab containing the antigen binding domain in accordance with SEQ ID NO: 21 and SEQ ID NO: 22.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, where the antibody competes with an antibody containing a CDR from mAb1-1371.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody contains the variable domains from mAb1-1371 in accordance with SEQ ID NO: 65 and SEQ ID NO: 66.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, wherein the antibody comprises a CDR from mAb1-1371 according to SEQ ID NO: 65 and SEQ ID NO: 66.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category", referred to herein as "category A", as a Fab containing the variable domains from mAb1-1371 in accordance with SEQ ID NO: 65 and SEQ ID NO: 66.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as mAb1-1371 in accordance with SEQ ID NO: 65 and SEQ ID NO: 66.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as the antibody or Fab containing the antigen binding domain in accordance with SEQ ID NO: 65 and SEQ ID NO: 66.

In one embodiment, an antibody of the present invention is capable of specifically binding FX/FXa, where the antibody competes with an antibody comprising a CDR from mAb1-1376, mAb1-6705, mAb1-7388, or mAb1-7563. Such antibodies are referred to herein as belonging to category B.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, wherein the antibody comprises the variable domains of mAb1-1376, mAb1-6705, mAb1-7388, or mAb1-7563, as defined by SEQ ID NOs: 67 and 68, SEQ ID NOs: 23 and 24, SEQ ID NOs: 39 and 40 and SEQ ID NOs: 59 and 60, respectively.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody contains the CDRs of mAb1-1376, mAb1-6705, mAb1-7388 or mAb1-7563, defined by SEQ ID NO: 67 and 68, SEQ ID NO: 23 and 24, SEQ ID NOs: 39 and 40 and SEQ ID NOs: 59 and 60, respectively.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as a Fab containing the variable domains of mAb1-1376, mAb1-6705, mAb1-7388, or mAb1-7563, defined according to SEQ ID NOs: 67 and 68, SEQ ID NOs: 23 and 24, SEQ ID NOs: 39 and 40, and SEQ ID NOs: 59 and 60, respectively.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as mAb1-1376, mAb1-6705, mAb1-7388 or mAb1-7563, as defined by SEQ ID NO: 67 and 68, SEQ ID NOs: 23 and 24, SEQ ID NOs: 39 and 40, and SEQ ID NOs: 59 and 60, respectively.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as the antibody or Fab containing the antigen binding domain in accordance with SEQ ID NO: 67 and 68, SEQ ID NO: 23 and 24, SEQ ID NO: 39 and 40 or SEQ ID NO: 59 and 60.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, where the antibody comprises CDRs or variable domains from an antibody selected from the group consisting of: mAb1-6723, 1-6716, 1-6721, 1-6730, 1-6731 , 1-6737, 1-6754, 1-7378, 1-7413, 1-7424, 1-7466, 1-7481, 1-7483 and mAb1-7591.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, where the antibody comprises CDRs or variable domains from an antibody selected from the group consisting of: mAb1-6723, 1-6716, 1-6721, 1-6730, 1-6731 , 1-6737, 1-6754, 1-7378, 1-7413, 1-7424, 1-7466, 1-7481, 1-7483, 1-7591, 1-7388, 1-7563, 1-7462 and mAb1 -7571.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as the mAb selected from the group of mAbs containing variable sequences or CDRs thereof selected from: SEQ ID NOs: 21 and 22 , SEQ ID NO: 25 and 26, SEQ ID NO: 27 and 28, SEQ ID NO: 29 and 30, SEQ ID NO: 31 and 32, SEQ ID NO: 33 and 34, SEQ ID NO: 35 and 36, SEQ ID NO: 37 and 38, SEQ ID NO: 39 and 40, SEQ ID NO: 41 and 42, SEQ ID NO: 43 and 44, SEQ ID NO: 45 and 46, SEQ ID NO: 51 and 52, SEQ ID NO : 53 and 54, SEQ ID NOs: 55 and 56, SEQ ID NOs: 57 and 58, SEQ ID NOs: 59 and 60, SEQ ID NOs: 61 and 62 and SEQ ID NOs: 63 and 64. This "category" of antibodies referred to herein as category C and is illustrated by a large number of individual antibodies, such as mAb1-6723, 1-6716, 1-6721, 1-6730, 1-6731, 1-6737, 1-6754, 1-7378, 1- 7413, 1-7424, 1-7466, 1-7481, 1-7483, 1-7591, 1-7388, 1-7563, 1-7462 and mAb1-7571.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, wherein the antibody competes for binding to FX, FX zymogen, or FXa with a reference antibody selected from the group of antibodies consisting of mAbs containing variable sequences or CDRs thereof selected from: SEQ ID NO: 21 and 22, SEQ ID NO: 25 and 26, SEQ ID NO: 27 and 28, SEQ ID NO: 29 and 30, SEQ ID NO: 31 and 32, SEQ ID NO: 33 and 34, SEQ ID NO : 35 and 36, SEQ ID NO: 37 and 38, SEQ ID NO: 41 and 42, SEQ ID NO: 43 and 44, SEQ ID NO: 53 and 54, SEQ ID NO: 55 and 56, SEQ ID NO: 57 and 58 and SEQ ID NO: 63 and 64.

In one embodiment, an antibody or antigen binding fragment thereof of the present invention competes for binding to FX/FXa with an antigen binding fragment comprising the CDRs of SEQ ID NOs: 21 and 22, SEQ ID NOs: 25 and 26, SEQ ID NOs: 27 and 28, SEQ ID NO: 29 and 30, SEQ ID NO: 31 and 32, SEQ ID NO: 33 and 34, SEQ ID NO: 35 and 36, SEQ ID NO: 37 and 38, SEQ ID NO: 41 and 42, SEQ ID NOs: 43 and 44, SEQ ID NOs: 53 and 54, SEQ ID NOs: 55 and 56, SEQ ID NOs: 57 and 58, or SEQ ID NOs: 63 and 64.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, where the antibody competes with an antibody comprising a CDR from mAb 1-7447, 1-7441, 1-7571 or 1-7462. They are referred to herein as category D antibodies.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody contains the variable domains of mAb1-7447, 1-7441, 1-7571 or 1-7462 in accordance with SEQ ID NO: 47 and 48, SEQ ID NO: 45 and 46, SEQ ID NOs: 51 and 53 or SEQ ID NOs: 61 and 62, respectively.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody contains the variable domains of mAb1-7447 or 1-7441 in accordance with SEQ ID NOs: 47 and 48, SEQ ID NOs: 45 and 46, respectively.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa, wherein the antibody comprises the CDRs of mAb1-7447, 1-7441, 1-7571, or 1-7462 according to SEQ ID NOs: 47 and 48, SEQ ID NO: 45 and 46, SEQ ID NOs: 51 and 53 or SEQ ID NOs: 61 and 62, respectively.

In one embodiment, an antibody of the present invention is capable of binding FX/FXa according to SEQ ID NO: 2, where the antibody belongs to the same "category" as a Fab containing the variable domains of mAb1-7447, 1-7441, 1- 7571 or mAb1-7462 according to SEQ ID NOs: 47 and 48, SEQ ID NOs: 45 and 46, SEQ ID NOs: 51 and 53 or SEQ ID NOs: 61 and 62, respectively.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as mAb1-7447, 1-7441, 1-7571 or mAb1-7462 in accordance with SEQ ID NO: 47 and 48, SEQ ID NOs: 45 and 46, SEQ ID NOs: 51 and 53 or SEQ ID NOs: 61 and 62, respectively.

In one embodiment, the antibody of the present invention is capable of binding FX/FXa, where the antibody belongs to the same "category" as the antibody or Fab containing the antigen binding domain in accordance with SEQ ID NOs: 47 and 48, SEQ ID NO: 45 and 46, SEQ ID NO: 51 and 53 or SEQ ID NO: 61 and 62.

In one embodiment, the antibody of the present invention is an antibody according to any of the preceding embodiments, wherein the antibody specifically binds the FX zymogen.

In such an embodiment, the antibody specifically binds the FX zymogen according to amino acid residues 1-139, 143-448 of SEQ ID NO: 2.

In one embodiment, the antibody of the present invention is an antibody according to any of the preceding embodiments, wherein the antibody binds FX.

In one embodiment, the antibody of the present invention is an antibody according to any of the preceding embodiments, wherein the antibody binds FXa.

In one such embodiment, the antibody specifically binds FXa according to amino acid residues 1-139, 195-448 of SEQ ID NO: 2.

In one embodiment, the antibody is a monospecific antibody. In one embodiment, the antibody is a multispecific antibody. In one such embodiment, the antibody is a bispecific antibody. In one such embodiment, the bispecific antibody is capable of binding to FIX or its activated form (FIXa) and FX/FXa. In one such embodiment, the bispecific antibody is capable of specifically binding to FIX/FIXa and FX/FXa.

In one embodiment, the antibody is a bispecific antibody capable of binding FIX/FIXa and FX/FXa, wherein the FIX/FIXa binding domain is derived from a Category 1 antibody and the FX/FXa binding domain is derived from a Category A antibody.

In one embodiment, the antibody is a bispecific FIX/FIXa and FX/FXa binding antibody, wherein the FIX/FIXa binding domain is derived from a Category 2 antibody and the FX/FXa binding domain is derived from a Category A antibody.

In one embodiment, the antibody is a bispecific FIX/FIXa and FX/FXa binding antibody, wherein the FIX/FIXa binding domain is derived from a Category 2 antibody and the FX/FXa binding domain is derived from a Category B antibody.

In one embodiment, the antibody is a bispecific FIX/FIXa and FX/FXa binding antibody, wherein the FIX/FIXa binding domain is derived from a Category 2 antibody and the FX/FXa binding domain is derived from a Category B antibody.

In one embodiment, the antibody is a bispecific FIX/FIXa and FX/FXa binding antibody, wherein the FIX/FIXa binding domain is derived from a category 1 antibody and the FX/FXa binding domain is derived from a category C or D antibody.

In one embodiment, the antibody is a bispecific antibody that binds FIX/FIXa and FX/FXa, where the binding domains are derived from mAb pairs consisting of: mAb1-1371/mAb1-1307, mAb1-6705/mAb1-1307, mAb1-1371/ mAb0-1886, mAb1-7441/mAb0-1886, mAb1-7447/mAb0-1886, mAb1-7481/mAb0-1886, mAb1-1371/mAb0-1998, mAb1-6716/mAb0-1998, mAb1-6723/mAb0 - 1998, mAb1-6730/mAb0-1998, mAb1-6731/mAb0-1998, mAb1-6737/mAb0-1998, mAb1-6754/mAb0-1998, mAb1-7378/mAb0-1998, mAb1-7441/mAb0-199 8, mAb1-7447/mAb0-1998, mAb1-7481/mAb0-1998, mAb1-1371/mAb1-4707, mAb1-6705/mAb1-4707, mAb1-1371/mAb1-4071, mAb1-7441/mAb1-5788, mAb1 - 7447/mAb1-5788, mAb1-7481/mAb1-5788, mAb1-1371/mAb1-4857, mAb1-6716/mAb1-4857, mAb1-6723/mAb1-4857, mAb1-6730/mAb1-4857, mAb1-673 1/ mAb1-4857, mAb1-6737/mAb1-4857, mAb1-6754/mAb1-4857, mAb1-7378/mAb1-4857, mAb1-7441/mAb1-4857, mAb1-7447/mAb1-4857 or mAb1-7481/mAb1 - 4857.

In one embodiment, the bispecific antibody of the present invention comprises an FX-binding antibody arm and a FIX/FIXa-binding antibody arm. In one such embodiment, the FX binding arm of the antibody binds to an epitope containing one or more residues from the FX activation peptide, and the FIX/FIXa binding arm of the antibody binds to an epitope containing one or more residues from the FIX protease domain.

In one embodiment, the antibody of the present invention is a multispecific antibody, such as a bi- or trispecific antibody.

In one embodiment, the antibody of the present invention is in an IgG format, such as full length IgG4.

In one embodiment, the antibody of the present invention is a chemical conjugate of two antibody fragments, such as a conjugate of two Fab fragments or scFv fragments or combinations thereof.

In one embodiment, the antibody of the present invention is a human or humanized antibody.

In one embodiment, the antibodies disclosed herein are intermediates for use in the manufacture of a bispecific antibody.

In one embodiment, the present invention includes antibodies that compete for binding to FIX/FIXa with the antibodies disclosed herein.

The antibody of the present invention can be used to treat a subject with coagulopathy and, in particular, with hemophilia A. Thus, the present invention also relates to the use of a monoclonal antibody that is capable of binding the FIX/FIXa protease domain for the treatment of a subject in need thereof; as well as the use of said antibody for the manufacture of a medicinal product for the treatment of a subject in need thereof. In addition, the present invention includes a method of treating a subject in need thereof with a monoclonal antibody that is capable of binding to the FIX/FIXa protease domain.

In one embodiment, an antibody of the present invention is capable of binding FIXa with a higher affinity than that with which it binds FIX.

In one embodiment, the antibody of the present invention is capable of increasing the enzymatic activity of FIXa towards FX.

In one such embodiment, an antibody of the present invention is capable of increasing the enzymatic activity of FIXa towards FX, as measured in an FXa production assay using the monovalent partial antibodies described herein.

In one embodiment, an antibody of the present invention is capable of increasing the enzymatic activity of FIXa towards FX, as measured in an FXa production assay using the bivalent antibodies described herein.

In one embodiment, the antibody of the present invention is not the anti-FIX antibody CLB-FIX 13, which is described in Rohlena et al. (2003) J Biol. Chem. 278(11):9394-9401. In one embodiment, the antibody of the present invention is not an anti-FIX HIX-1 (mouse IgG1) antibody (Merck KGaA, SigmaAldrich). In one embodiment, the antibody of the present invention is not an anti-FIX antibody AHIX-5041 (IgG1) (Haematologic Technologies, Inc.).

In one embodiment, the antibody of the present invention has reduced immunogenicity compared to procoagulant antibodies of the prior art.

In one embodiment, the bispecific antibody or antigen binding fragment thereof comprises a first antigen binding region recognizing FIX (SEQ ID NO: 1) or an activated form thereof (FIXa) and a second antigen binding region recognizing FX (SEQ ID NO: 2) or an activated form thereof (FXa), where

a) the first antigen binding region contains the following CDR sequences:

V _H -CDR1: DYAMH

V _H -CDR2: GISWRGDIIGYVDSVKG

V _H -CDR3: SYGSGSFYNAFDS

V _L -CDR1: RASQSISSWLA

V _L -CDR2: KASRLDR

V _L -CDR3: LEYSSYIRT

And

b) the second antigen binding region contains the following CDR sequences:

_VH -CDR1: TSWIV

V _H -CDR2: MIDPSDSFTSYSPSFQG

V _H -CDR3: LHYYHSEEFDV

V _L -CDR1: RASQSVSSSYLA

V _L -CDR2: GASSRAR

V _L -CDR3: QQFGSSRLFT

In one embodiment, the bispecific antibody or antigen binding fragment thereof comprises a first antigen binding region recognizing FIX (SEQ ID NO: 1) or an activated form thereof (FIXa) and a second antigen binding region recognizing FX (SEQ ID NO: 2) or an activated form thereof (FXa), where

a) the first antigen binding region contains the following CDR sequences:

V _H -CDR1: DYAMH

V _H -CDR2: GISWRGDIIGYVDSVKG

V _H -CDR3: SYGSGSFYNAFDS

V _L -CDR1: RASQSISSWLA

V _L -CDR2: KASRLDR

V _L -CDR3: LEYSSYIRT

And

b) the second antigen binding region contains the following CDR sequences:

_VH -CDRI: TSWIV

V _H -CDR2: MIDPSDSFTSYSPSFQG

V _H -CDR3: LHYYHSEEFDV

V _L -CDR1: RASQSVSSSYLA

V _L -CDR2: GASSRTR

V _L -CDR3: QQFGSSRLFT

The present invention is further described by the following embodiments.

1. An antibody or antigen-binding fragment thereof capable of binding to factor IX (FIX) in accordance with SEQ ID NO: 1 or its activated form (FIXa).

2. The antibody or antigen binding fragment thereof of Embodiment 1, wherein the antibody or antigen binding fragment thereof is part of "category 1".

3. The antibody or antigen binding fragment thereof of Embodiment 1, wherein the antibody or antigen binding fragment thereof competes with a reference antibody, wherein the reference antibody comprises

a. a heavy chain variable domain identified by SEQ ID NO: 15 and a light chain variable domain identified by SEQ ID NO: 16, or

b. a heavy chain variable domain identified under SEQ ID NO: 19, and a light chain variable domain identified under SEQ ID NO: 20.

4. The antibody of the preceding embodiment, wherein the reference antibody is a Fab.

5. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises

A. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 15, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 16,

b. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 19, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 20,

c. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 69, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 70,

d. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 71, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 72,

e. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 73, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 74,

f. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 83, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 84,

g. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 81, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 82,

h. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 75, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 76,

i. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 77, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 78, or

j. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 177, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 178.

6. The antibody or antigen binding fragment thereof of Embodiment 5, wherein the heavy chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

7. The antibody or antigen binding fragment thereof of Embodiment 5, wherein the light chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

8. The antibody or antigen binding fragment thereof of embodiments 6 and 7, wherein both the heavy chain variable domain and the light chain variable domain are at least 92%, 94%, 96% or 98% identical to the identified SEQ IDs.

9. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises

A.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 15, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 16,

b.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 19, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 20,

With.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 69, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 70,

d.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 71, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 72,

e.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 73, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 74,

f.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 83, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 84,

g.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 81, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 82,

h.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 75, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 76, or

i.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 77, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 78, or

j

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 177, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 178.

10. The antibody or antigen binding fragment thereof of Embodiment 9, wherein the three heavy chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

11. The antibody or antigen binding fragment thereof of Embodiment 9, wherein the three heavy chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

12. The antibody or antigen binding fragment thereof of Embodiment 9, wherein the three light chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

13. The antibody or antigen binding fragment thereof of Embodiment 9, wherein the three light chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

14. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 15 and the light chain variable domain CDR sequence identified under SEQ ID NO: 16, or

b. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 19 and the light chain variable domain CDR sequence identified under SEQ ID NO: 20, or

c. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 3 and the light chain variable domain CDR sequence identified under SEQ ID NO: 4, or

d. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 109 and the light chain variable domain CDR sequence identified under SEQ ID NO: 110, or

e. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 153 and the light chain variable domain CDR sequence identified under SEQ ID NO: 154, or

f. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 171 and the light chain variable domain CDR sequence identified under SEQ ID NO: 172, or

g. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 177 and the light chain variable domain CDR sequence identified under SEQ ID NO: 178, or

h. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 187 and the light chain variable domain CDR sequence identified under SEQ ID NO: 188.

15. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. a heavy chain variable domain identified by SEQ ID NO: 15 and a light chain variable domain identified by SEQ ID NO: 16, or

b. a heavy chain variable domain identified by SEQ ID NO: 19 and a light chain variable domain identified by SEQ ID NO: 20, or

c. a heavy chain variable domain identified by SEQ ID NO: 3 and a light chain variable domain identified by SEQ ID NO: 4, or

d. the heavy chain variable domain identified by SEQ ID NO: 109 and the light chain variable domain identified by SEQ ID NO: 110, or

e. a heavy chain variable domain identified under SEQ ID NO: 153 and a light chain variable domain identified under SEQ ID NO: 154, or

f. a heavy chain variable domain identified under SEQ ID NO: 171 and a light chain variable domain identified under SEQ ID NO: 172, or

g. a heavy chain variable domain identified under SEQ ID NO: 177 and a light chain variable domain identified under SEQ ID NO: 178, or

h. a heavy chain variable domain identified under SEQ ID NO: 187, and a light chain variable domain identified under SEQ ID NO: 188.

16. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising one or more amino acid residues of L337, R338, S339, T340, K341 and T343 of SEQ ID NO: 1.

17. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residue R338 of SEQ ID NO: 1.

18. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residues R338 and K341 of SEQ ID NO: 1.

19. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising two or three amino acid residues of L337, R338, S339, T340, K341 and T343 of SEQ ID NO: 1.

20. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising four or five amino acid residues of L337, R338, S339, T340, K341 and T343 of SEQ ID NO: 1.

21. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing

a. R338, S339, T340, K341 and T343,

b. L337, S339, T340, K341 and T343,

c. L337, R338, T340, K341 and T343,

d. L337, R338, S339, K341 and T343,

e. L337, R338, S339, T340 and T343 or

f. L337, R338, S339, T340 and K341

from SEQ ID NO: 1.

22. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residues L337, R338, S339, T340, K341 and T343 of SEQ ID NO: 1.

23. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising one or more amino acid residues of K301, D332, R333, A334, T335, R338 and N346 of SEQ ID NO: 1 .

24. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residues L337, R338, S339, T340, K341 and T343 of SEQ ID NO: 1.

25. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising two or three amino acid residues of K301, D332, R333, A334, T335, R338 and N346 of SEQ ID NO: 1 .

26. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising four or five amino acid residues of K301, D332, R333, A334, T335, R338 and N346 of SEQ ID NO: 1 .

27. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising five or six amino acid residues of K301, D332, R333, A334, T335, R338 and N346 of SEQ ID NO: 1 .

28. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing

a. D332, R333, A334, T335, R338 and N346,

b. K301, R333, A334, T335, R338 and N346,

c. K301, D332, A334, T335, R338 and N346,

d. K301, D332, R333, T335, R338 and N346,

e. K301, D332, R333, A334, R338 and N346,

f. K301, D332, R333, A334, T335 and N346 or

g. K301, D332, R333, A334, T335 hR338

from SEQ ID NO: 1.

29. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residues D332, R333, L337 and R338 of SEQ ID NO: 1.

30. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residues K301, D332, R333, A334, T335, R338 and N346 of SEQ ID NO: 1.

31. The antibody or antigen-binding fragment thereof of Embodiment 1, wherein the antibody or antigen-binding fragment thereof belongs to "category 2".

32. The antibody or antigen binding fragment thereof of Embodiment 1, wherein the antibody or antigen binding fragment thereof competes with a reference antibody, wherein the reference antibody comprises a heavy chain variable domain identified by the sequence of SEQ ID NO: 17 and a light chain variable domain identified by sequences under SEQ ID NO: 18.

33. The antibody of the preceding embodiment, wherein the reference antibody is a Fab.

34. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 17, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 18,

b. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 85, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 86, or

c. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 79, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 80.

35. The antibody or antigen binding fragment thereof of embodiment 34, wherein the heavy chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

36. The antibody or antigen binding fragment thereof of embodiment 34, wherein the light chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

37. The antibody or antigen binding fragment thereof of embodiments 35 and 36, wherein both the heavy chain variable domain and the light chain variable domain are at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

38. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

A.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 17, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 18,

b.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 85, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 86, or

With.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 79, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 80.

39. The antibody or antigen binding fragment thereof of embodiment 38, wherein the three heavy chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

40. The antibody or antigen binding fragment thereof of embodiment 38, wherein the three heavy chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

41. The antibody or antigen binding fragment thereof of Embodiment 38, wherein the three light chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

42. The antibody or antigen binding fragment thereof of embodiment 38, wherein the three light chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

43. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

A. the CDR sequence of the heavy chain variable domain identified under SEQ ID NO: 17, and

the CDR sequence of the light chain variable domain identified under SEQ ID NO: 18.

44. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

A. a heavy chain variable domain identified under SEQ ID NO: 17, and a light chain variable domain identified under SEQ ID NO: 18.

45. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising one or more amino acid residues of H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411 from SEQ ID NO: 1.

46. The antibody or antigen-binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing amino acid residues H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411 of SEQ ID NO : 1.

47. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing two, three or four amino acid residues of H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411 from SEQ ID NO: 1.

48. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing five, six or seven amino acid residues of H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411 from SEQ ID NO: 1.

49. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing eight, nine or ten amino acid residues of H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411 from SEQ ID NO: 1.

50. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing amino acid residues

a. H257, N258, K293, R403, Y404, N406, W407, E410 and K411,

b. H256, N258, K293, R403, Y404, N406, W407, E410 and K411,

c. H256, H257, K293, R403, Y404, N406, W407, E410 and K411,

d. H256, H257, N258, R403, Y404, N406, W407, E410 and K411,

e. H256, H257, N258, K293, Y404, N406, W407, E410 and K411,

f. H256, H257, N258, K293, R403, N406, W407, E410 and K411,

g. H256, H257, N258, K293, R403, Y404, W407, E410 and K411,

h. H256, H257, N258, K293, R403, Y404, N406, E410 and K411,

i. H256, H257, N258, K293, R403, Y404, N406, W407 and K411 or

j. H256, H257, N258, K293, R403, Y404, N406, W407 and E410 from SEQ ID NO: 1.

51. The antibody or antigen-binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing amino acid residues H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411 of SEQ ID NO : 1.

52. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residues H257, K293 and N406 of SEQ ID NO: 1.

53. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody is a procoagulant antibody.

54. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of increasing the procoagulant activity of FIXa.

55. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody is capable of increasing the enzymatic activity of FIXa towards FX.

56. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody is capable of functionally replacing FVIII and/or FVIIIa.

57. An antibody or antigen-binding fragment thereof capable of binding to FX (SEQ ID NO: 2) or its activated form (FXa).

58. The antibody or antigen binding fragment thereof of embodiment 57, wherein the antibody or antigen binding fragment thereof is part of "category A".

59. The antibody or antigen binding fragment thereof of embodiment 57, wherein the antibody or antigen binding fragment thereof competes with a reference antibody, wherein the reference antibody comprises a heavy chain variable domain identified under SEQ ID NO: 65 and a light chain variable domain identified under SEQ ID NO: 66.

60. The antibody of the preceding embodiment, wherein the reference antibody is a Fab.

61. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing one or more amino acid residues of H101, E103, R113, T116, L117, A118, T127, S227, E228, F229 , Y230, E266, R287, L303, P304, E305, L419, K420, D423, R424, M426, K427 and T428 from FX/FXa.

62. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is capable of binding an epitope containing amino acid residues H101, E103, R113, T116, L117, A118, T127, S227, E228, F229, Y230, E266 , R287, P304, L303, P304, E305, L419, K420, D423, R424, M426, K427 and T428 from FX/FXa.

63. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising one or more amino acid residues of R113, Y230, K420, D423, R424 and K427 of FX/FXa.

64. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is capable of binding an epitope comprising amino acid residues R113, Y230, K420, D423, R424 and K427 of FX/FXa.

65. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain at least 90% identical to the sequence identified in SEQ ID NO: 65 and a light chain variable domain of at least 90% identical to the sequence identified under SEQ ID NO: 66.

66. The antibody or antigen binding fragment thereof of embodiment 65, wherein the heavy chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

67. The antibody or antigen binding fragment thereof of embodiment 65, wherein the light chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

68. The antibody or antigen binding fragment thereof of embodiments 66 and 67, wherein both the heavy chain variable domain and the light chain variable domain are at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

69. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 65, and

b. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 66.

70. The antibody or antigen binding fragment thereof of Embodiment 69, wherein the three heavy chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

71. The antibody or antigen binding fragment thereof of Embodiment 69, wherein the three heavy chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

72. The antibody or antigen binding fragment thereof of Embodiment 69, wherein the three light chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

73. The antibody or antigen binding fragment thereof of Embodiment 69, wherein the three light chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

74. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises the heavy chain variable domain CDR sequences identified under SEQ ID NO: 65 and the light chain variable domain CDR sequences identified under SEQ ID NO: 66.

75. The antibody or antigen binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable domain identified under SEQ ID NO: 65 and a light chain variable domain identified under SEQ ID NO: 66.

76. The antibody or antigen binding fragment thereof of embodiment 57, wherein the antibody or antigen binding fragment thereof is part of "category B".

77. The antibody or antigen-binding fragment thereof of Embodiment 57, wherein the antibody or antigen-binding fragment thereof competes with a reference antibody, wherein the reference antibody comprises a heavy chain variable domain identified by SEQ ID NO: 67 and a light chain variable domain identified by SEQ ID NO: 68.

78. The antibody or antigen binding fragment thereof of the preceding embodiment, wherein the reference antibody is a Fab.

79. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 67, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 68,

b. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 23, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 24,

c. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 39, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 40, or

d. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 59, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 60.

80. The antibody or antigen binding fragment thereof of embodiment 79, wherein the heavy chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

81. The antibody or antigen binding fragment thereof of embodiment 79, wherein the light chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

82. The antibody or antigen binding fragment thereof of embodiments 80 and 81, wherein both the heavy chain variable domain and the light chain variable domain are at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

83. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

A.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 67, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 68,

b.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 23, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 24,

With.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 39, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 40, or d.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 59, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 60.

84. The antibody or antigen binding fragment thereof of Embodiment 83, wherein the three heavy chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs of the identified SEQ ID.

85. The antibody or antigen binding fragment thereof of embodiment 83, wherein the three heavy chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

86. The antibody or antigen binding fragment thereof of Embodiment 83, wherein the three light chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

87. The antibody or antigen binding fragment thereof of Embodiment 83, wherein the three light chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

88. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises

a. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 67, and the light chain variable domain CDR sequence identified under SEQ ID NO: 68,

b. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 23, and the light chain variable domain CDR sequence identified under SEQ ID NO: 24,

c. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 39 and the light chain variable domain CDR sequence identified under SEQ ID NO: 40, or

d. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 59 and the light chain variable domain CDR sequence identified under SEQ ID NO: 60.

89. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises

a. a heavy chain variable domain identified under SEQ ID NO: 67, and a light chain variable domain identified under SEQ ID NO: 68,

b. a heavy chain variable domain identified under SEQ ID NO: 23, and a light chain variable domain identified under SEQ ID NO: 24,

c. a heavy chain variable domain identified by SEQ ID NO: 39 and a light chain variable domain identified by SEQ ID NO: 40, or

d. a heavy chain variable domain identified under SEQ ID NO: 59, and a light chain variable domain identified under SEQ ID NO: 60.

90. The antibody or antigen binding fragment thereof of embodiment 57, wherein the antibody or antigen binding fragment thereof is part of "category C".

91. The antibody or antigen binding fragment thereof of Embodiment 57, wherein the antibody or antigen binding fragment thereof competes with a reference antibody, wherein the reference antibody comprises a heavy chain variable domain identified by SEQ ID NO: 21 and a light chain variable domain identified by SEQ ID NO: 22.

92. The antibody or antigen binding fragment thereof of the preceding embodiment, wherein the reference antibody is a Fab.

93. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 21, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 22,

b. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 25, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 26,

c. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 27, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 28,

d. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 29, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 30,

e. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 31, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 32,

f. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 33, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 34,

g. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 35, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 36,

h. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 37, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 38,

i. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 39, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 40,

j. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 41, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 42,

k. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 43, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 44,

l. a heavy chain variable domain at least 90% identical to the sequence, a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 51, and a light chain variable domain at least 90% identical to the sequence, identified under SEQ ID NO: 52,

m. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 53, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 54,

n. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 55, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 56,

O. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 57, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 58,

R. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 59, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 60,

q. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 61, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 62, or

r. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 63, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 64.

94. The antibody or antigen binding fragment thereof of embodiment 93, wherein the heavy chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

95. The antibody or antigen binding fragment thereof of embodiment 93, wherein the light chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

96. The antibody or antigen binding fragment thereof of embodiments 94 and 95, wherein both the heavy chain variable domain and the light chain variable domain are at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

97. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises

A.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 21, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 22,

b.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 25, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 26,

With.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 27, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 28, or

d.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 29, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 30,

e.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 31, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 32,

f.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 33, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 34,

g.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 35, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 36, or

h.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 37, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 38,

i.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 39, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 40,

j.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 41, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 42,

k.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 43, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 44, or

l.

m.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 51, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 52, or

n.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 53, and

three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 54,

O.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 55, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 56,

p.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 57, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 58,

q.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 59, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 60,

r.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 61, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 62, or

s.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 63, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 64.

98. The antibody or antigen binding fragment thereof of Embodiment 97, wherein the three heavy chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

99. The antibody or antigen binding fragment thereof of embodiment 97, wherein the three heavy chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

100. The antibody or antigen binding fragment thereof of Embodiment 97, wherein the three light chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs of the identified SEQ ID.

101. The antibody or antigen binding fragment thereof of Embodiment 97, wherein the three light chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

102. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 21, and the light chain variable domain CDR sequence identified under SEQ ID NO: 22,

b. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 25, and the light chain variable domain CDR sequence identified under SEQ ID NO: 26,

c. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 27, and the light chain variable domain CDR sequence identified under SEQ ID NO: 28,

d. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 29, and the light chain variable domain CDR sequence identified under SEQ ID NO: 30,

e. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 31, and the light chain variable domain CDR sequence identified under SEQ ID NO: 32,

f. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 33, and the light chain variable domain CDR sequence identified under SEQ ID NO: 34,

g. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 35, and the light chain variable domain CDR sequence identified under SEQ ID NO: 36,

h. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 37, and the light chain variable domain CDR sequence identified under SEQ ID NO: 38,

i. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 39, and the light chain variable domain CDR sequence identified under SEQ ID NO: 40,

j. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 41, and the light chain variable domain CDR sequence identified under SEQ ID NO: 42,

k. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 43, and the light chain variable domain CDR sequence identified under SEQ ID NO: 44,

l. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 51, and the light chain variable domain CDR sequence identified under SEQ ID NO: 52,

m. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 53, and the light chain variable domain CDR sequence identified under SEQ ID NO: 54,

n. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 55, and the light chain variable domain CDR sequence identified under SEQ ID NO: 56,

O. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 57, and the light chain variable domain CDR sequence identified under SEQ ID NO: 58,

R. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 59, and the light chain variable domain CDR sequence identified under SEQ ID NO: 60,

q. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 61 and the light chain variable domain CDR sequence identified under SEQ ID NO: 62, or

r. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 63, and the light chain variable domain CDR sequence identified under SEQ ID NO: 64,

103. An antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable domain and a light chain variable domain identified by

a. SEQ ID NO: 21 and 22,

b. SEQ ID NO: 25 and 26,

c. SEQ ID NO: 27 and 28,

d. SEQ ID NO: 29 and 30,

e. SEQ ID NO: 31 and 32,

f. SEQ ID NO: 33 and 34,

g. SEQ ID NO: 35 and 36,

h. SEQ ID NO: 37 and 38,

i. SEQ ID NO: 39 and 40,

j. SEQ ID NO: 41 and 42,

k. SEQ ID NO: 43 and 44,

l. SEQ ID NO: 51 and 52,

m. SEQ ID NO: 53 and 54,

n. SEQ ID NO: 55 and 56,

O. SEQ ID NO: 57 and 58,

p. SEQ ID NO: 59 and 60,

q. SEQ ID NO: 61 and 62 or

r. SEQ ID NO: 63 and 64 respectively.

104. The antibody or antigen-binding fragment thereof of embodiment 57, wherein the antibody or antigen-binding fragment thereof is "category D".

105. The antibody or antigen binding fragment thereof of Embodiment 57, wherein the antibody or antigen binding fragment thereof competes with a reference antibody, wherein the reference antibody comprises a heavy chain variable domain identified by SEQ ID NO: 47 and a light chain variable domain identified by SEQ ID NO: 48.

106. The antibody of the preceding embodiment, wherein the reference antibody is a Fab.

107. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 47, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 48,

b. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 45, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 46,

c. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 51, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 52,

or

d. a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 61, and a light chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 62.

108. The antibody or antigen binding fragment thereof of embodiment 107, wherein the heavy chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

109. The antibody or antigen binding fragment thereof of embodiment 107, wherein the light chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

110. The antibody or antigen binding fragment thereof of embodiments 108 and 109, wherein both the heavy chain variable domain and the light chain variable domain are at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

111. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

A.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 47, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 48,

b.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 45, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 46,

With.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 51, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 52, or

d.

i. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 61, and

ii. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 62.

112. The antibody or antigen binding fragment thereof of embodiment 111, wherein the three heavy chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

113. The antibody or antigen binding fragment thereof of embodiment 111, wherein the three heavy chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

114. The antibody or antigen binding fragment thereof of embodiment 111, wherein the three light chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs of the identified SEQ ID.

115. The antibody or antigen binding fragment thereof of embodiment 111, wherein the three light chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

116. An antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 47, and the light chain variable domain CDR sequence identified under SEQ ID NO: 48,

b. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 45, and the light chain variable domain CDR sequence identified under SEQ ID NO: 46,

c. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 51, and the light chain variable domain CDR sequence identified under SEQ ID NO: 52,

or

d. the heavy chain variable domain CDR sequence identified under SEQ ID NO: 61, and the light chain variable domain CDR sequence identified under SEQ ID NO: 62.

117. An antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. a heavy chain variable domain identified under SEQ ID NO: 47, and a light chain variable domain identified under SEQ ID NO: 48,

b. a heavy chain variable domain identified under SEQ ID NO: 45, and a light chain variable domain identified under SEQ ID NO: 46,

c. a heavy chain variable domain identified under SEQ ID NO: 51, and a light chain variable domain identified under SEQ ID NO: 52,

or

d. a heavy chain variable domain identified under SEQ ID NO: 61, and a light chain variable domain identified under SEQ ID NO: 62.

118. The antibody or antigen-binding fragment thereof of Embodiment 57, wherein the antibody or antigen-binding fragment thereof is "category E".

119. The antibody or antigen binding fragment thereof of Embodiment 57, wherein the antibody or antigen binding fragment thereof competes with a reference antibody, wherein the reference antibody comprises a heavy chain variable domain identified by SEQ ID NO: 49 and a light chain variable domain identified by SEQ ID NO: 50.

120. The antibody of the preceding embodiment, wherein the reference antibody is a Fab.

121. The antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain at least 90% identical to the sequence identified under SEQ ID NO: 49 and a light chain variable domain of at least 90% identical to the sequence identified under SEQ ID NO: 50.

122. The antibody or antigen binding fragment thereof of embodiment 121, wherein the heavy chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ ID.

123. The antibody or antigen binding fragment thereof of embodiment 121, wherein the light chain variable domain is at least 92%, 94%, 96%, or 98% identical to the identified SEQ ID.

124. The antibody or antigen binding fragment thereof of embodiments 122 and 123, wherein both the heavy chain variable domain and the light chain variable domain are at least 92%, 94%, 96%, or 98% identical to the identified SEQ IDs.

125. An antibody or antigen-binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises

a. three heavy chain CDR sequences with no more than 10 amino acid substitutions compared to the heavy chain variable domain CDR sequences identified under SEQ ID NO: 49, and

b. three light chain CDR sequences with no more than 10 amino acid substitutions compared to the light chain variable domain CDR sequences identified under SEQ ID NO: 50.

126. The antibody or antigen binding fragment thereof of embodiment 125, wherein the three heavy chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs identified SEQ ID.

127. The antibody or antigen binding fragment thereof of embodiment 125, wherein the three heavy chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

128. The antibody or antigen binding fragment thereof of embodiment 125, wherein the three light chain CDR sequences have no more than 9, such as 8, such as 7, or such as 6, amino acid substitutions compared to the CDRs of the identified SEQ ID.

129. The antibody or antigen binding fragment thereof of embodiment 125, wherein the three light chain CDR sequences have no more than 5, such as 4, such as 3, such as 2, or no more than 1 amino acid substitution at compared to the CDRs from the identified SEQ IDs.

130. The antibody or antigen binding fragment thereof according to any of the preceding embodiments, wherein the antibody or antigen binding fragment thereof comprises the heavy chain variable domain CDR sequences identified under SEQ ID NO: 49 and the light chain variable domain CDR sequences identified under SEQ ID NO: 50.

131. The antibody or antigen-binding fragment thereof according to any one of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain identified under SEQ ID NO: 49 and a light chain variable domain identified under SEQ ID NO: 50.

132. Polyspecific antibody or antigen-binding fragment thereof, capable of binding to FIX/FIXa and FX/FXa.

133. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 1-131.

134. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 2-30.

135. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 31-52.

136. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 57-131.

137. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 58-75.

138. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 76-89.

139. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 90-103.

140. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 104-117.

141. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 118-131

142. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 1-56 and the antigen binding fragment of any of the preceding embodiments 57-131.

143. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 2-30 and the antigen binding fragment of any of the preceding embodiments 58-75.

144. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 2-30 and the antigen binding fragment of any of the preceding embodiments 76-89.

145. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 2-30 and the antigen binding fragment of any of the preceding embodiments 90-103.

146. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 31-52, the antigen binding fragment of any of the preceding embodiments 58-75.

147. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 31-52 and the antigen binding fragment of any of the preceding embodiments 76-89.

148. The polyspecific antibody or antigen binding fragment thereof of embodiment 132, wherein the antibody comprises the antigen binding fragment of any of the preceding embodiments 31-52 and the antigen binding fragment of any of the preceding embodiments 90-103.

149. The polyspecific antibody or antigen-binding fragment thereof of embodiment 132, wherein the antibody is a bispecific antibody capable of specifically binding FIX/FIXa and FX/FXa, wherein the binding domains are derived from mAb pairs consisting of: mAb1-1371/mAb1-1307, mAb1-6705/mAb1-1307, mAb1-1371/mAb0-1886, mAb1-7441/mAb0-1886, mAb1-7447/mAb0-1886, mAb1-7481/mAb0-1886, mAb1-1371/mAb0-1998, mAb1 - 6716/mAb0-1998, mAb1-6723/mAb0-1998, mAb1-6730/mAb0-1998, mAb1-6731/mAb0-1998, mAb1-6737/mAb0-1998, mAb1-6754/mAb0-1998, mAb1-737 8/ MAB0-1998, MAB1-7441/MAB0-1998, MAB1-7447/MAB0-1998, Mab/Mab0-1998, Mab1-6723/MAB1-1307, MAB1-6723/MAB0-1886, MAB1-6705/MAB0-1886, mAb1-7481/mAb0-1998 and mAb1-6705/mAb0-1998.

150. The antibody or antigen-binding fragment thereof of any one of embodiments 132-149, wherein the antibody is a procoagulant bispecific antibody.

151. The antibody or antigen binding fragment thereof as in any one of embodiments 132-149, wherein the antibody is a bispecific antibody capable of enhancing the procoagulant activity of FIXa.

152. The antibody or antigen binding fragment thereof as in any one of embodiments 132-149, wherein the antibody is a bispecific antibody capable of increasing the enzymatic activity of FIXa towards FX.

153. The antibody or antigen-binding fragment thereof of any one of embodiments 132-149, wherein the antibody is a bispecific antibody capable of functionally replacing FVIII and/or FVIIIa.

154. A polyspecific antibody capable of stimulating the enzymatic activity of FIXa towards FX, containing a first antigen binding region recognizing FIX (SEQ ID NO: 1) or its activated form (FIXa), and a second antigen binding region recognizing FX (SEQ ID NO: 2 ) or its activated form (FXa), where

a) the first antigen-binding region contains a CDR from an antibody selected from the group consisting of: mAb1-5743, mAb1-6584, mAb1-8768, mAb1-6037, mAb1-6081, mAb1-4857, mAb1-8780, mAb1-9016, mAb1 -9015, mAb1-8467, mAb1-5783, or mAb1-5781, and

b) the second antigen binding region comprises a CDR from an antibody selected from the group consisting of: mAb1-6738, mAb1-6463, mAb1-6723, mAb1-7503 or mAb1-6097.

155. Polyspecific antibody capable of stimulating the enzymatic activity of FIXa towards FX, containing

the first polypeptide recognizing FIX/FIXa, and

a second polypeptide recognizing FX/FXa, where

a) the first polypeptide contains a heavy chain variable domain and a light chain variable domain from mAb1-5743, mAb1-6584, mAb1-8768, mAb1-6037, mAb1-6081, mAb1-4857, mAb1-8780, mAb1-9016, mAb1-9015 , mAb1-8467, mAb1-5783 or mAb1-5781, and

b) the second polypeptide contains a heavy chain variable domain and a light chain variable domain from mAb1-6738, mAb1-6463, mAb1-6723, mAb1-7503 or mAb1-6097.

156. The antibody of any one of embodiments 154 or 155, wherein the antibody is a bispecific antibody.

157. The antibody of any one of embodiments 150-156, wherein stimulation of FIXa enzymatic activity toward FX is determined in the FXa production assay described herein using a monovalent partial anti-FIX/FIXa antibody, wherein the stimulation index is at least 94, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 22 00, 2300, 2400, 2500-fold, 3000-fold, 4000-fold, or 5000-fold when measured using a partial antibody concentration resulting in FIXa saturation of at least 80%.

158. Polyspecific antibody capable of stimulating the enzymatic activity of FIXa towards FX, containing

a first antigen binding region recognizing FIX (SEQ ID NO: 1) or an activated form thereof (FIXa), and a second antigen binding region recognizing FX (SEQ ID NO: 2) or an activated form thereof (FXa), where

the first antigen binding site is capable of specifically binding an epitope containing amino acid residue R338 of FIX/FIXa, and

wherein the second antigen binding site is capable of binding to the EGF-2 domain and/or the FX/FXa catalytic subunit.

159. The polyspecific antibody of embodiment 158, wherein

the first antigen-binding site is capable of specifically binding an epitope containing amino acid residues R338 and K341 of FIX/FIXa.

160. The polyspecific antibody of embodiment 159, wherein

the first antigen-binding site is capable of specifically binding an epitope containing amino acid residues L337, R338, S339, T340, K341 and T343 of FIX/FIXa.

161. A polyspecific antibody capable of stimulating the enzymatic activity of FIXa towards FX, containing a first antigen binding region recognizing FIX (SEQ ID NO: 1) or its activated form FIXa, and a second antigen binding region recognizing FX (SEQ ID NO: 2) or its activated form (FXa),

Where

the first antigen binding site is capable of specifically binding an epitope comprising amino acid residues D332, R333, L337 and R338 of FIX/FIXa, and

wherein the second antigen binding site is capable of binding to the EGF-2 domain and/or the FX/FXa catalytic subunit.

162. The polyspecific antibody of embodiment 161, wherein

the first antigen-binding site is capable of specifically binding an epitope containing amino acid residues K301, D332, R333, A334, T335, R338, N346 from FIX/FIXa.

163. Polyspecific antibody capable of stimulating the enzymatic activity of FIXa towards FX, containing

a first antigen binding site recognizing FIX/FIXa, and

a second antigen-binding site recognizing FX/FXa, where

the first antigen-binding site is capable of specifically binding an epitope containing amino acid residues H257, K293 and N406 of FIX/FIXa, and

wherein the second antigen binding site is capable of binding to the EGF-2 domain and/or the FX/FXa catalytic subunit.

164. The antibody of any one of embodiments 1-56 and 132-156, wherein stimulation of FIXa enzymatic activity toward FX is measured in the FXa production assay described herein using a monovalent partial anti-FIX/FIXa antibody, wherein the stimulation index is at least 94, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100 , 2200, 2300-fold, 2400-fold, 2500-fold, 3000-fold, 4000-fold, or 5000-fold when measured using a partial antibody concentration resulting in FIXa saturation of at least 80%.

165. The antibody of any one of embodiments 1-56 and 132-156, wherein stimulation of FIXa enzymatic activity toward FX is measured in the FXa production assay described herein using a monovalent partial anti-FIX/FIXa antibody, wherein the stimulation index is 50-5000 times, such as 94-2500 times, such as 100-2500 times, such as 200-2500 times, such as 300-2500 times, such as , 400-2500-fold, such as 500-2500-fold, such as 600-2500-fold, such as 700-2500-fold, such as 800-2500-fold, such as , 500-2500-fold, such as 600-2500-fold, such as 700-2500-fold, such as 800-2500-fold, such as 900-2500-fold, such as , 1000-2500-fold, such as 1500-2500-fold, when measured using a partial antibody concentration resulting in FIXa saturation of at least 80%.

166. The antibody of any one of the preceding embodiments, wherein the antibody is a procoagulant antibody.

167. An antibody according to any of the preceding embodiments, wherein said antibody functionally replaces FVIII and/or FVIIIa.

168. The antibody of any one of the preceding embodiments, wherein the antibody is a bispecific antibody.

169. The antibody of any one of the preceding embodiments, wherein the isotype of the antibody is IgG1, IgG2, IgG3, or IgG4, or a combination thereof.

170. The antibody of any one of embodiments 57-131, wherein the light chain variable domain of said antibody or antigen binding fragment thereof comprises amino acid residues R57, R96 (SEQ ID NO: 22) and wherein the heavy chain variable domain of said antibody comprises amino acid residues W33, D52 , D55, H100, Y101, Y102, H103 (SEQ ID NO: 21).

171. An antibody or antigen-binding fragment thereof according to any of the preceding embodiments for use in a method of treating a coagulopathy or bleeding disorder.

172. A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof according to any of the preceding embodiments for the treatment of a coagulopathy or bleeding disorder.

173. A method of treating a subject suffering from a coagulopathy or bleeding disorder, comprising administering to said subject an antibody or antigen binding fragment thereof according to any of the preceding embodiments.

174. The method of embodiment 173, wherein the coagulopathy or bleeding disorder is hemophilia A or hemophilia A in the presence of inhibitors.

EXAMPLES

Example 1. Development of expression plasmids for Fab and mAb to factor IX/FIXa

The FIX/FIXa binding antibodies disclosed herein have been identified using various antibody development methods. To create a diverse set of antibodies targeting FIXa and FX, immunization of mice and rabbits, as well as selection from phage display and Adimab yeast display, were performed.

Adimab Yeast Display

The Adimab platform is a yeast display system containing a fully human naïve IgG1/kappa library with 10 ¹⁰ diversity covering 20 of 42 VH families. The antibody phage display platform used is a proprietary, fully human Fab display library. The library is 10 ^{× 10} in size and was generated by a combination approach using chemical synthesis of the light chain and heavy chain CDR1 and CDR2, complemented by PCR amplification of the heavy chain CDR3 from human peripheral blood mononuclear cells. The antibody selection process is controlled using MACS and FACS based methods which allow monitoring of the applied selection criteria in real time. Since selection procedures are based on MACS and FACS, labeled antigens (eg, biotin) are required. Selection operations were performed using biotin-labeled active site inhibited hFIXa (FIXa-EGR-biotin) or antibody-mediated immobilization of hFIXa. The hits were assessed for binding using biolayer interferometry (ForteBio Octet system).

Phage display

To maximize epitope diversity coverage, various panning strategies were explored, including panning using biotinylated FIXa-EGR, FX, active site inhibited FXa, or antigen capture using anti-FIXa antibodies. Initial hits were identified using phage ELISA. Following sequence analysis, unique hits were cloned, expressed as IgG1, and ranked using SPR (Biacore) or biolayer interferometry (ForteBio Octet system).

In vivo platforms

Kymouse™ HK and HL 1.0 mice (producing kappa and lambda chains, respectively) were used to generate fully human antibodies in mice. Additional immunization procedures were performed in wild-type mice and rabbits to maximize antibody diversity.

Obtaining antibodies to FIXa

Mice or rabbits were immunized with FIXa, FIXa-EGR, or FX using standard protocols. Antibodies produced in mice or rabbits were screened using ELISA. Rabbit FIXa-binding B cells were sorted by FACS using randomly biotinylated FIXa-EGR. Antibody “hits” from rabbits and mice were either recombinantly expressed (rabbit mAbs) or expanded (mouse hybridomas) and the antibodies were subsequently purified on a small scale.

Obtaining antibodies to FX

Kymouse mice and rabbits were immunized with FX using standard or repeat immunization multiple site (RIMMS) protocols. Rabbit B cells were isolated by FACS sorting using random biotinylated FX. Anti-FX mAbs from fusions and sorts were screened using ELISA and ForteBio Octet systems.

Sequencing of antibodies obtained from Kymouse mice and wt mice

Anti-FIXa and anti-FX antibodies produced by hybridomas derived from Kymouse mice or wt mice were sequenced and expressed in HEK293 cells using standard techniques. Expressed antibodies were assessed for binding using ForteBio Octet systems.

The resulting DNA sequences encoding the variable domains ( _VH and _VL ) of the selected antibodies were introduced into a pTT-based mammalian expression vector (Durocher et al (2002) Nucleic Acid Res. 30:E9) or into a pcDNA3.4 mammalian expression vector (Invitrogen ), containing DNA sequences encoding the antibody constant regions. For the pTT/pcDNA3.4 mAb expression vectors, the DNA sequences for V _H and V _L were inserted in frame with DNA sequences encoding the constant regions of human IgG1 or IgG ₄ S228P ( _CH 1C _H 2C _H 3) or human C _L kappa chain respectively. For the corresponding Fab expression vectors pTT/pcDNA3.4, the DNA sequences for V _H were brought in frame with the DNA sequences encoding human IgG _{1 CH} 1.

For the reference compound 224F3 used in Examples 6, 8 and 18 below, the _VH and _VL sequences of 224F3 were obtained according to EP 1660536 B1 (SEQ ID NO: 1 and 2, respectively). DNA sequences encoding V _H and V _L 224F3 were introduced into a mammalian expression vector based on pTT5/pcDNA3.4 in reading frame with DNA sequences encoding the human IgG1 constant region ( _CH 1C _H 2C _H 3) or C _L kappa chain person accordingly.

All expression vectors included a 5' DNA sequence containing a kozak sequence and a DNA sequence encoding the signal peptide in frame with the DNA sequences encoding the antibody.

Example 2. Recombinant expression of antibodies and Fab fragments of antibodies

Antibodies and antibody Fab fragments were expressed using transient transfection of HEK293 cell suspension (293Expi, Invitrogen), essentially following the manufacturer's instructions. 293Expi cells were routinely subcultured every 3-4 days in Expi293F expression medium (Invitrogen, cat. no. A1435104) supplemented with 1% P/S (GIBCO, cat. no. 15140-122). Expi293F cells were transfected at a cell density of 2.5-3 million/ml using Expifectamine. For each liter of Expi293F cells, transfection was performed by diluting a total of 1 mg of plasmid DNA (V _{H -CH} 1 (for Fab) or V _{H -CH} 1-C _H 2-C _H 3 (for mAb) and LC plasmids in a 1:1 ratio) in 50 ml of Optimem (GIBCO, catalog number 51985-026, dilution A) and by diluting 2.7 ml of Expifectamine in 50 ml of Optimem (dilution B). For joint transfections with the production of Fab and mAb, V _H -C _H 1 and LC plasmids (Fab) and V _H -C _H 1-C _H 2-C _H 3 and LC plasmids (mAb) were used, respectively, in a ratio of 1 :1. Dilutions A and B were mixed and incubated at room temperature for 10-20 minutes. The transfection mixture was then added to Expi293F cells and the cells were incubated at 37°C in a humidified incubator with epicyclic rotation (85-125 rpm). One day after transfection, 5 ml of ExpiFectamine 293 transfection enhancer 1 and 50 ml of ExpiFectamine 293 transfection enhancer 2 were added to transfected cells. Cell culture supernatants were typically collected 4-5 days after transfection by centrifugation followed by filtration.

Example 3 Purification and Characterization of Fab and Antibodies

Fab Purification and Characterization

Purification of Fab molecules was performed as a 2-step process consisting of affinity chromatography using kappaSelect resin (GE Healthcare, part number 17-5458-11) and size exclusion chromatography using Superdex200 resin (GE Healthcare, part number 17-1043-11). 04). Purification procedures were carried out using a chromatographic system (GE Healthcare part number 18-1112-41). The buffer systems used for the affinity purification step were an equilibration buffer consisting of 20 mM Na phosphate, pH 7.2, 150 mM NaCl, and an elution buffer consisting of 10 mM formic acid, pH 3.5, and a pH adjustment buffer , consisting of 0.4 M Na phosphate, pH 9.0. Cell supernatants were applied directly without any adjustments to a pre-equilibrated kappaSelect SuRe column. The column was washed with 10 column volumes of equilibration buffer and Fab molecules were eluted isocratically in approximately 5 column volumes of elution buffer. The pH of the collected fractions was adjusted to neutral using the described pH adjustment buffer immediately after elution. Fab molecules were further purified and buffer exchanged using the specified gel filtration resin prepacked on the column. The running buffer used for size exclusion chromatography was 25 mM HEPES and 150 mM NaCl, pH 7.4. Fab molecules eluted as discrete peaks at approximately 0.5 column volume. Peak fractions were analyzed using size exclusion high performance liquid chromatography (SE-HPLC) on an Agilent LC 1100/1200 system using a BIOSep-SEC-S3000 300×7.8 mm column (Phenomenex, catalog number 00H-2146- K0) and a working buffer consisting of 200 mM Na phosphate, pH 6.9, 300 mM NaCl and 10% isopropanol. Based on this analysis, the fractions were combined to obtain a homogeneous protein preparation. The final drug eluted as a single symmetrical peak with a retention time of approximately 10 min. at a flow rate of 1 ml/min.

Purified Fab molecules were further characterized using SDS-PAGE/Coomassie and liquid chromatography-mass spectrometry analyses. SDS-PAGE/Coomassie analysis was performed using 4-12% NuPage Bis-Tris gels (Invitrogen, cat. no. NP0321BOX). All Fab molecules showed the expected light chain and heavy chain components. Intact molecular weight determination procedures were performed using a liquid chromatography-electrospray ionization time-of-flight mass spectrometry setup on an Agilent 6210 instrument and a MassPREP desalting column (Waters, catalog number USRM10008656). The buffer system used was an equilibration buffer consisting of LC-MS grade 0.1% formic acid in H ₂ O and an elution buffer consisting of LC-MS grade 0.1% formic acid in ACN. All Fab molecules showed the expected intact molecular weight values according to the sequence. Final purity was determined based on SE-HPLC analysis. All purity estimates were between 95-99% for the various Fab fragments. To determine final protein concentrations, absorbance was measured at 280 nm using a NanoDrop spectrophotometer (Thermo Scientific) and concentrations were calculated using specific extinction coefficients for each of the Fab molecules.

Antibody Purification and Characterization

Antibody purification was performed by affinity chromatography using MabSelect SuRe Protein A Resin (GE Healthcare, cat. no. 17-5438-01). Purification procedures were carried out using a chromatographic system (GE Healthcare part number 18-1112-41). The buffer systems used for the affinity purification step were an equilibration buffer, which consisted of 20 mM Na phosphate, pH 7.2, 150 mM NaCl, and an elution buffer, consisting of 10 mM formic acid, pH 3.5, and an adjustment buffer pH consisting of 0.4 M Na phosphate, pH 9.0. Cell supernatants were applied directly without any adjustments to a pre-equilibrated MabSelect SuRe column. The column was washed with 10 column volumes of equilibration buffer and the antibodies were eluted isocratically in approximately 2-5 column volumes of elution buffer. The pH of the collected fractions was adjusted to neutral using the described pH adjustment buffer immediately after elution.

Purified antibodies were characterized using SDS-PAGE/Coomassie, size exclusion liquid chromatography (SE-HPLC) and liquid chromatography-mass spectrometry (LC-MS) analyses. SDS-PAGE/Coomassie analysis was performed using 4-12% NuPage Bis-Tris gels (Invitrogen, cat. no. NP0321BOX). In this case, all antibodies showed the expected light chain and heavy chain components. Intact molecular weight determination procedures were performed using a liquid chromatography-electrospray ionization time-of-flight mass spectrometry setup on an Agilent 6210 instrument and a MassPREP desalting column (Waters, catalog number USRM10008656). The buffer system used was an equilibration buffer consisting of LC-MS grade 0.1% formic acid in H ₂ O and an elution buffer consisting of LC-MS grade 0.1% formic acid in ACN. Assays were performed with and without N-glycosidase F (Roche Diagnostics, catalog number 11365177001) and a reducing agent (i.e., mercaptoethanol or DTT). All antibodies showed the expected intact molecular weight values according to the sequence and one heavy chain N-glycan. Purity was determined based on SE-HPLC. The final protein purity was analyzed based on the SE-HPLC method setup on an Agilent LC 1100/1200 system and using a BIOSep-SEC-S3000 300×7.8 mm column (Phenomenex, catalog number 00H-2146-K0) and a running buffer consisting of from 200 mM Na phosphate, pH 6.9, 300 mM NaCl and 10% isopropanol. UV280 and fluorescence detectors (exc. 280 nm/emission 354 nm) were used for detection. Antibodies eluted as single symmetrical peaks with retention times reflecting antibody size. All purity estimates were between 95-99% for different antibodies. A NanoDrop spectrophotometer (Thermo Scientific) was used to measure final protein concentrations along with specific extinction coefficients for each antibody.

Example 4. Categorization of stimulating antibodies to FIXa

Antibodies selected as capable of stimulating the enzymatic activity of FIXa towards FX were analyzed in categorization experiments to determine the binding characteristics of the identified antibodies using the method described below.

Antibody categorization method

Categorization experiments were performed using ForteBio Octet systems (HTX, Red384) equipped with anti-human IgG sensors (Pall Life Sciences, Menlo Park, CA) and using 8- or 32-channel mode (Red384 and HTX). Categorization analyzes were performed using a classic sandwich epitope categorization setup. Briefly, (1) the first antibody was captured with anti-human ANS tips (anti-human IgG Fc capture tips (ANS, cat. no. 18-5064), (2) unblocked IgG binding sites on the ANS tips were blocked with human polyclonal IgG (14506 SIGMA ), (3) FIXa was bound to the primary antibody, (4) a competing antibody was offered to the antibody-antigen complex on the tips, and if secondary antibody binding could not be detected, then the antibodies were scored as belonging to the same category.

The analysis identified two different categories, category 1 and 2, defined by antibodies mAb0-1886 and mAb1-1307, respectively.

Categorization of antibodies to FIX.

The selected FIX antibodies were categorized against each other and two different categories were identified (category 1 and 2). The numbers refer to the mAb ID, for example 0-1998 indicates mAb0-1998.

The review shows that several antibodies were classified as category 1, including mAb0-1886 and mAb0-1998. Category 2 was found to include four antibodies in addition to mAb1-1307. Two antibodies, mAb1-0072 and mAb1-0073, were common to categories 1 and 2.

The parent antibody variants (lineages) disclosed herein fall into the same categories and share epitope residues (hot spots) with the parent antibodies.

Because the antibody variants for which data are provided in this Example do not contain amino acid substitutions at positions that are critical for epitope recognition, as demonstrated by the crystal structures of the parent antibody-FIXa complexes presented in Example 5, one skilled in the art should be aware It is clear that the starting point variants will belong to the same category, compete for binding and recognize at least the same hot spot residues in the FIX/FIXa epitope as the antibody from which they originate, i.e. mAb0-1998, mAb0-1886 or mAb1-1307.

Example 5: Crystallization and Epitope Mapping of Anti-FIX/FIXa Antibodies Using X-Ray Crystallography

The FIXa protein used for crystallization (Cambridge ProteinWorks, product code 10316) consists of a truncated light chain (residues 85-142 of SEQ ID NO: 1) with a non-natural methionine residue attached at the N-terminus as a result of bacterial expression, and a heavy chain , containing residues 181-415 of SEQ ID NO: 1. The active site of the protease is blocked with EGR-chloromethyl ketone.

Crystallization

Fab0-7237:FIXa

Crystals of Fab0-7237 (Fab fragment corresponding to mAb0-1886), mixed in a 1:1 molar ratio with the FIXa protein, were grown using the hanging drop vapor diffusion technique at 18°C. A solution of 0.8 μl of 7.5 mg/ml protein in 20 mM Tris-HCl, pH 7.4, 50 mM NaCl and 2.5 mM CaCl ₂ was mixed with an equal volume of 4 M sodium formate as a precipitant and incubated in 1 ml precipitator.

Fab0-7238:FIXa

Crystals of Fab0-7238 (Fab fragment corresponding to mAb0-1998), mixed in a 1:1 molar ratio with the FIXa protein, were grown using the sessile drop vapor diffusion technique at 18°C. A protein solution of 0.1 μl of 6.2 mg/ml in 20 mM Tris-HCl, pH 7.4, 50 mM NaCl and 2.5 mM CaCl ₂ was mixed with 0.1 μl of 100 mM sodium cacodylate, pH 6.5, and 1 M trisodium citrate as a precipitant and incubated with 60 μl of precipitant.

Fab0-7236:FIXa

Crystals of Fab0-7236 (Fab fragment corresponding to mAb 1-1307), mixed in a 1:1 molar ratio with the FIXa protein, were grown using the sessile drop vapor diffusion technique at 18°C. A solution of 0.1 μl of 6.4 mg/ml protein in 20 mM Tris-HCl, pH 7.4, 50 mM NaCl and 2.5 mM CaCl ₂ was mixed with an equal volume of 0.2 M lithium sulfate, 40% (vol. /vol.) PEG400 and 0.1 M Tris, pH 8.5, as a precipitant and incubated with 1 ml of precipitant.

Diffraction Data Collection

Fab0-7237:FIXa

The crystal was cryoprotected in a solution consisting of 3 M sodium formate, 4% glycerol, 4% ethylene glycol, 4.5% sucrose and 1% glucose before flash cooling in liquid nitrogen. Diffraction data were collected at 100K in the X06DA synchrotron radiation beam from Swiss Light Source (wavelength 1.0000 ) using the Pilatus2M pixel detector from Dectris. Auto-indexing, integration and scaling of data were performed using programs from the XDS package (statistical characteristics of diffraction data are summarized in Table 1).

Fab0-7238:FIXa

The crystal was cryoprotected in a solution consisting of 75 mM sodium cacodylate, pH 6.5, and 0.75 M trisodium citrate, 4% glycerol, 4% ethylene glycol, 4.5% sucrose, and 1% glucose before flash cooling in liquid nitrogen. Diffraction data were collected at 100K in the X06DA synchrotron radiation beam from Swiss Light Source (wavelength 1.0000 ) using the Pilatus2M pixel detector from Dectris. Auto-indexing, integration and scaling of data were performed using programs from the XDS package (statistical characteristics of diffraction data are summarized in Table 1).

Fab0-7236:FIXa

Three crystals were cryoprotected in a solution consisting of 0.15 M lithium sulfate, 30% (v/v) PEG400 and 0.075 M Tris, pH 8.5, 4% glycerol, 4% ethylene glycol, 4.5% sucrose and 1% glucose before flash cooling in liquid nitrogen. Diffraction data were collected at 100K in the X06DA synchrotron radiation beam from Swiss Light Source (wavelength 1.0000 ) using the Pilatus2M pixel detector from Dectris. Auto-indexing, integration, merging and scaling of data were performed using programs from the XDS package (statistical characteristics of diffraction data are summarized in Table 1).

Defining and clarifying the structure

Fab0-7237:FIXa

The structure was determined by molecular replacement using Phaser implemented in the Phenix software suite, with the H and L chains from Protein Structure Database entry 4NP4 and the H and L chains from Protein Structure Database entry 3KCG. The asymmetric unit contains two Fab:FIXa complexes. The model was refined using refinement steps in Phenix and manual rebuilding in COOT. Statistical characteristics of the refinement are shown in Table 1.

Fab0-7238:FIXa

The structure was determined by molecular replacement using Phaser implemented in the Phenix software suite, with the H and L chains from the Protein Structure Database entry 4PUB and the H and L chains from the Protein Structure Database entry 3KCG. The asymmetric unit contains two Fab:FIXa complexes. The model was refined using refinement steps in Phenix and manual rebuilding in COOT. Statistical characteristics of the refinement are shown in Table 1.

Fab0-7236:FIXa

The structure was determined by molecular replacement using Phaser implemented in the Phenix software suite, the Fab portion of the complex structure of the Fab0-7238:FIXa complex described above, and the H and L chains from the Protein Structure Database entry 3KCG. The asymmetric unit contains one Fab:FIXa complex. The model was refined using refinement steps in Phenix and manual rebuilding in COOT. Statistical characteristics of the refinement are shown in Table 1.

Statistical characteristics for the highest resolution shell are shown in parentheses.

Determination of epitopes

Based on the above mAb0-1998, mAb1-1307 and mAb0-1886 were found to bind to different epitopes on FIXa, where an epitope is defined as residues having at least one heavy atom within a distance of ≤ 3.5 from the heavy atom in the antibody.

The mAb0-1998 epitope is located in the 170-loop and contains the following protease domain residues: L337, R338, S339, T340, K341 and T343.

The mAb1-1307 epitope contains the following residues: H256, H257, N258, K293, R403, Y404, N406, W407, E410 and K411.

The mAb0-1886 epitope is located in a 170-helix and contains the following residues of the protease domain: K301, D332, R333, A334, T335, R338 and N346.

The epitopes of mAb0-1998 and mAb0-1886 were found to overlap, which is in good agreement with the observation that these two antibodies compete for binding to FIX/FIXa (Example 4).

The parent antibody variants (lineages) disclosed herein fall into the same categories and share epitope residues (hot spots) with the parent antibodies.

Because the antibody variants reported in Example 4 above and in certain Examples below do not contain amino acid substitutions at positions that are critical for epitope recognition as demonstrated by the crystal structures of the parent antibody-FIXa complexes presented in this Example, one skilled in the art will appreciate that the variants will, as a starting point, be of the same category, compete for binding, and recognize at least the same hot spot residues in the FIX/FIXa epitope as the antibody from which they are derived. occur, i.e. mAb0-1998, mAb0-1886 or mAb1-1307.

Example 6: Activity of bivalent antibodies to FIX/FIXa in the FXa production assay

The stimulatory effect of bivalent anti-FIX/FIXa antibodies on the enzymatic activity of FIXa towards FX was determined by their ability to stimulate activation of FX by FIXa in the presence of a procoagulant phospholipid membrane according to the principles described in Scheiflinger et al. (2008) J Thromb Haemost, 6:315–322. Given the high anti-FIXa activity of antibody 224F3 among the antibodies identified by Scheiflinger et al., 224F3 was chosen as the reference in the following experiments (see Example 1 for design information on 224F3).

The stimulatory effect of anti-FIXa antibodies on FIXa-mediated activation of FX in FXa was measured in an automated high-throughput biochemical assay in 384-well plates. Briefly, FIXa was mixed with purified antibody at a four-point 5-fold dilution for dose response. The FX/phospholipid (PL) mixture was added and the extent of FXa formation upon addition of FXa substrate (Pefaflour) was measured and the rate of substrate hydrolysis was determined by detecting fluorescence for five minutes on a multi-label reader (PheraSTAR). The relative stimulatory activity of FIXa was calculated as the rate of formation of FXa with the FIXa-antibody complex compared with FIXa alone.

Each antibody was tested over a concentration range of 0–200 nM by preincubation with 3 nM human plasma-derived FIXa (Haematologic Technologies Inc, USA) and 10 μM phospholipid vesicles of a 25:75 phosphatidylserine:phosphatidylcholine mixture (Haematologic Technologies Inc, USA) in assay buffer (50 mM HEPES, 100 mM NaCl, 5 mM CaCl ₂ , 0.1% (wt/vol) PEG8000, pH 7.3, +1 mg/ml BSA) for 10 min. before adding FX obtained from human blood plasma (Haematologic Technologies Inc, USA) to a concentration of 150 nM. After 10 min activation at room temperature, the reaction (50 μl) was quenched by adding 25 μl quenching buffer (50 mM HEPES, 100 mM NaCl, 60 mM EDTA, 0.1% PEG8000, pH 7.3, +1 mg/ml BSA ). The amount of FXa produced was then determined by adding 25 μL of 2 mM chromogenic substrate S-2765 (Chromogenix, Sweden) and measuring the conversion of the chromogenic substrate by measuring the optical density at 405 im (ΔOD/min) in a microplate reader. The rate of FXa formation at each antibody concentration was determined using a standard curve constructed with known amounts of FXa obtained from human plasma (Haematologic Technologies Inc, USA).

Table 2 shows the measured rates of FXa production for each antibody at the concentrations tested. From this, peak stimulatory activity values were calculated for each antibody as the observed maximum rate of FXa production compared to the rate of production with 224F3. These data are presented in Table 3, which shows that antibodies belonging to each of the three families (0-1886, 0-1998 and 1-1307, respectively) have activities that are 10-67 times higher than the activity observed for 224F3 (Scheiflinger et al.).

Example 7. Preparation of monovalent (partial) antibodies

To avoid any possible avidity effects associated with conventional monospecific and bivalent antibodies, such as in FXa production assays (Example 8) and certain SPR experiments (Examples 14 and 15), a monovalent partial (OA) antibody format was used as described by Martens et al.: A Novel One-Armed Anti-c-Met Antibody Inhibits Glioblastoma Growth In Vivo. Clin. Cancer Res. 12, 6144-6152 (2006), where the full heavy chain, the truncated heavy chain (without the Fab region) and the light chain are expressed together. Instead of co-expression of the three chains described by Martens et al., the present invention produced monovalent antibodies using the ^Duobody® principle as described for bispecific antibodies (Example 10). Thus, monovalent antibodies were prepared by mixing the full monospecific and bivalent antibodies and a truncated heavy chain dimer (formally prepared from the full antibody by removing the Fab region) and allowing chains to be exchanged under the same experimental conditions as described in Example 10. To form a monovalent antibodies require that the antibody and the truncated heavy chain dimer carry the appropriate complementary mutations to promote heterodimerization, i.e. F405L/K409R for IgG1 and F405L+R409K/WT for IgG4, as described in example 10.

In the case of monovalent antibodies of the IgG1 subtype, heavy chain truncation can occur from the N-terminus to a position between Cys 220 and the upper hinge Cys 226 (EU numbering). A specific example of a truncated IgG1 heavy chain is one where residues 1-220 are truncated.

In the case of monovalent antibodies of the IgG4 subtype, heavy chain truncation can occur from the N-terminus to a position between Cys 200 and the upper hinge Cys 226 (EU numbering). A specific example of a truncated IgG4 heavy chain is one where residues 1-214 are truncated.

Example 8: Activity of monovalent antibodies to FIX/FIXa in the FXa production assay

To avoid any possible avidity effects arising from the bivalence of the standard antibody format, the stimulatory activity of anti-FIX/FIXa antibodies relative to the enzymatic activity of FIXa against FX was determined after conversion to a monovalent partial (OA) antibody format (see Example 8). The antibodies tested are listed in Table 4 below. A monovalent OA variant of the anti-FIXa antibody 224F3 (designated mAb1-1582), also mentioned in Example 7, was included for comparison.

OA antibody stimulatory activity was measured in assay buffer (50 mM HEPES, 100 mM NaCl, 5 mM CaCl ₂ , 0.1% (w/v) PEG8000, pH 7.3, +1 mg/mL BSA) at fixed concentrations phospholipid vesicles of a mixture of phosphatidylserine (PS): phosphatidylcholine (PC) (final concentration 500 μM; Haematologic Technologies Inc, USA) and plasma-derived FIXa (final concentrations 0.17, 0.5 or 1 nM; Haematologic Technologies Inc, USA) . The concentration of FIXa was chosen to ensure that less than 15% of the FX substrate was converted to FXa. After preincubation in the presence of monovalent OA antibody (final concentrations are listed in Table 1), 150 nM plasma-derived FX was added to produce a final reaction volume of 50 μl and activation was allowed to proceed for 20 min. at room temperature. The reaction was then quenched by adding 25 μl of quenching buffer (50 mM HEPES, 100 mM NaCl, 60 mM EDTA, 0.1% PEG8000, pH 7.3, +1 mg/ml BSA) and the amount of FXa produced was determined by further addition of 25 μl 2 mM chromogenic substrate S-2765 (Chromogenix, Sweden) and measuring the conversion of the chromogenic substrate by measuring the optical density at 405 nm (ΔOD/min) in a microplate reader. The measured activity was corrected for background activity by subtracting the signal measured in the same assay but replacing FIXa and antibody with assay buffer, and then normalized according to the concentration of FIXa present in the assay ([FIXa] _total ). By dividing this number by the similarly normalized rate of FXa formation in the absence of the antibody (A _FIXa,norm. ), the antibody stimulation index was calculated, providing multiple stimulation of FIXa activity by the antibody at the concentration used. Due to the low rate of FXa formation by free FIXa, activation reactions in the absence of antibody were performed as described above, but in the presence of 5, 10, or 20 nM FIXa. The measured activity values were then background subtracted and normalized according to the concentration of FIXa in the assay. The average of three normalized free FIXa activity values was used to calculate the stimulation index.

Determination of the stimulation index

Thus, the calculation of the stimulation index can be described as follows.

Stimulation index = ((A _FIXa+OA - A _background ) / FIXa] _general ) / A _{FIXa, normal.} ,

where A _FIXa+OA is the activity measured in the presence of the OA antibody, A _background is the background activity measured in the absence of FIXa and monovalent antibody, [FIXa] _total is the concentration of FIXa in the assay, and A _FIXa,norm. represents the mean of normalized free FIXa activity.

Determination of saturation FIXa

The proportion of FIXa saturated with OA antibody in the assay is determined from the concentrations of FIXa and OA antibody and the equilibrium dissociation constant (K _d ) governing their interaction. The latter can be measured using techniques known in the art, such as isothermal titrapion calorimetry (ITC).

Because the stimulation index will increase as the concentration of OA antibody increases until FIXa saturation is achieved, the concentration of OA antibody in the assay should be selected to ensure at least 80% saturation of FIXa in the assay to ensure proper assessment of the stimulation index at full saturation FIXa.

The fraction of FIXa bound to OA antibody at steady state (f _FIXa+OA ) can be calculated from the total concentrations of FIXa ([FIXa] _total ) and OA ([OA] _total ) in the assay and the equilibrium dissociation constant ( _Kd ) for them interactions using a quadratic coupling equation as described by Krishnaswamy et al. (1992) J. Biol. Chem., 267:23696-23706 and detailed in equations 1 and 2 below, where

[FIXa+OA] _{of the assay} represents the calculated concentration of the FIXa-OA-antibody complex at steady state in the assay,

f _FIXa+OA represents the calculated percentage of FIXa that is bound to the OA antibody at steady state in the assay.

The stimulation index for each monovalent OA antibody is shown in Table 4. For all antibodies tested, the stimulation index measured was found to be higher than the stimulation index measured for the monovalent partial antibody 224F3 (mAb1-1582).

At a concentration of partial antibody 224F3 in the assay of 3260 nM and a K _d for interaction with FIXa of 0.477 nM, as reported by Kerschbaumer et al. (US7297336-B2), more than 95% of FIXa was associated with the partial antibody 224F3 in the assay.

Example 9 Development of expression plasmids for Fab and mAb to FX/FXa

The anti-FX/FXa antibodies disclosed herein were developed using standard antibody development methods and expression plasmids were prepared as described in Example 1 for the expression plasmids for Fab and anti-FIX/FIXa mAbs. Expression, purification and characterization of anti-FX/Xa antibodies were also performed as described for anti-FIX/FIXa antibodies in Examples 2 and 3.

Example 10: Bispecific Antibodies Prepared by In Vitro Assembly

Bispecific antibodies are prepared by in vitro assembly of first and second antibodies using the Duobody ^® (Genmab) method described (Labrijn et al. PNAS 2013, vol. 110, pp. 5145-5150) for IgG1 bispecific antibodies, and using a slightly modified version for bispecific IgG4 antibodies, as detailed below.

For IgG1, the heavy chain constant region of the first antibody is K409R IgG1 (anti-FIX/FIXa), and the heavy chain constant region of the second antibody is F405L IgG1 (anti-FX/FXa). IgG1 may be a variant of IgG1 with reduced effector functions, as mentioned previously.

For IgG4, the heavy chain constant region of the first antibody is S228P IgG4 (anti-FIX/FIXa), and the heavy chain constant region of the second antibody is S228P F405L R409K IgG4 (anti-FX). Two parent antibodies are prepared as described in Examples 1-3. The Fab arm exchange reaction is carried out in HEPES buffer (pH 7.4) under reducing conditions using 75 mM 2-mercaptoethylamine (2-MEA) and incubation at 30°C for 3 hours.

Example 11. Procoagulant activity of bispecific antibodies

Anti-FIXa and anti-hFX antibody pairs were converted into bispecific antibodies using ^Duobody® technology as described above (Example 10). Bispecific antibodies were tested for procoagulant activity in various assays, such as the FXa generation assays described above (Example 6) and the thrombin generation test (TGT) described in the next paragraph.

Thrombin Generation Test (TGT) Analysis

TGT was performed in an automated 384-well HTP setup using kaolin stimulation (Haemonetics Corporation, catalog number 6300). Briefly, antibodies were added at a concentration of 111 nM (except for mAb1-1371, which was added at 55 nM, and mAb1-0021, which was added at 166 nM) to hemophilia A (HA) blood plasma (George King). Kaolin mixed with phospholipids (Rossix, part number PL604T) was then added, followed by the addition of FIIa substrate (FluCa, Thrombinoscope, part number TS50.00). Fluorescence was measured on an En Vision Perkin Elmer multilabel plate reader at 1-minute intervals for 2 hours. Peak height was calculated as the maximum value observed in the thrombogram and then normalized to the peak height observed for the reference anti-FIXa and anti-FX antibodies. The reference always included the binding domain from the anti-FX antibody mAb1-2375 (identified by SEQ ID NOs: 93 and 94) in combination with FIX domains from each of the three families represented by mAb1-4707, mAb1-5788 and mAb1-4857. Antibodies were grouped by their relative activity in TGT as low (0-24%), + (24-50%), ++ (50-75%) and +++ (>75%), with + being preferred, ++ is preferred, and +++ is most preferred.

Selection of preferred anti-FIXa/anti-FX bispecific antibody combinations

A large number of anti-FX antibodies were tested as bispecific antibodies in combination with anti-FIXa antibody variants belonging to three lineages, mAb1-1307, mAb0-1886 and mAb0-1998. Selected combinations of anti-FIXa antibody/anti-FX antibody pairs demonstrating significant activity in the TGT assay are shown in Table 5.

As can be seen from Table 5, the level of activity exhibited by the bispecific antibody depends on the specific anti-FIXa antibody/anti-FX antibody combination. For example, the anti-FX antibody mAb1-6723 in combination with the anti-FIXa antibody mAb0-1998 exhibits strong activity (+++), while the activity of mAb1-6723 in combination with mAb0-1886 (+) and mAb1-1307 (+) is lower.

Example 12: Categorization of Antibodies to FX

Some anti-FX antibodies exhibiting significant activity in TGT in a bispecific antibody format in combination with an anti-hFIXa antibody (Example 11) were categorized against each other using ForteBio Octet systems according to the same procedure as described for anti-FIXa antibodies (Example 4). , with the exception of replacing FIXa with FX.

The analysis identified five different categories, categories A-E, defined by antibodies mAb1-1371, mAb1-1376, mAb1-6723, mAb1-7447 and mAb1-7449, respectively. Two categories, category A and category E, are represented by only one anti-FX antibody (see Table 6).

A large number of clones were identified in category C as competitors of mAb1-6723.

Example 13 Crystallization and epitope mapping of an anti-FX antibody using X-ray crystallography

Crystallization

Attempts to crystallize Fab0-8954 (Fab fragment corresponding to mAb1-6723) in complex with FX were unsuccessful, while good quality crystals were obtained with FXa upon active site inhibition. Thus, Fab0-8954 crystals mixed in a 1:1 molar ratio with FXa without gla with inhibited active site (bacterially expressed human factor Xa without gla domain (wild type), inhibited EGR, lot #hGDFXAEGR-022, Cambridge Protein Works) , were grown using the vapor diffusion technique in the sessile drop version at 18°C. 150 nl of a protein solution of 6.7 mg/ml of the complex in 20 mM Tris-HCl, pH 7.4, 50 mM NaCl and 2.5 mM CaCl ₂ were mixed with 50 nl of 0.2 M magnesium acetate, 0.1 M sodium cacodylate , pH 6.5, 20% (wt/vol) PEG 8000 as precipitant and incubated with 60 μl of precipitant.

Diffraction Data Collection

The crystal was cryoprotected by adding 1 μL of precipitant with 20% ethylene glycol to the crystallization drop before flash cooling in liquid nitrogen. Diffraction data were collected at 100K in the X06DA synchrotron radiation beam from Swiss Light Source (wavelength 1.0000 ) using the Pilatus2M pixel detector from Dectris. Auto-indexing, integration and scaling of data were performed using programs from the XDS package (statistical characteristics of diffraction data are summarized in Table 7).

Defining and clarifying the structure

The asymmetric unit contains four Fab:FXa complexes, as inferred from Matthews coefficient analysis. The structure was determined by molecular replacement. Phaser, implemented in the Phenix software suite, was used with the H and L chains from the Protein Structure Database entry 5I1K as a search model localizing the four Fabs. These were modeled using the correct amino acid sequence using COOT and then refined using refinement in Phenix. The refined Fab model was refined using molecular replacement in Molrep from the CCP4 set with chains A and B from Protein Structure Database entry 1G2L as a search model. Four FXa fragments were discovered. The model was refined using refinement steps in Phenix and manual rebuilding in COOT. Statistical characteristics of the refinement are shown in Table 7.

Statistical characteristics for the highest resolution shell are shown in parentheses.

Epitope definition

The crystal structure of the Fab0-8954:FXa complex contains four copies of the complex in an asymmetric unit, and these were analyzed separately for epitope and paratope identification using a cutoff distance of 3.5 .

Residues are included in the epitope for mAb1-6723 if they satisfy the distance criterion of 3.5 in at least one of the four copies of the Fab0-8954:FXa complex in the unit cell. The epitope and paratope residues for mAb1-6723 are listed in Table 8.

The parent antibody variants (lineages) disclosed herein fall into the same categories and share epitope residues (hot spots) with the parent antibodies.

Because the antibody variants exemplified herein do not contain amino acid substitutions at positions that are critical for epitope recognition, as demonstrated by the crystal structures of the parent antibody-FXa complexes presented in this example, one of ordinary skill in the art It should be clear that the starting point variants will belong to the same category, compete for binding and recognize at least the same hot spot residues in the FX/FXa epitope as the antibody from which they originate, i.e. e. mAb1-6723.

Example 14: Identification of Hot Spot Remnants on FX

Similar to the mapping of epitope hot spot residues on FIX for mAb1-1307, mAb0-1886 and mAb0-1998 as described in Example 15, the data presented in this example defines epitope hot spot residues on FX for mAb1-6723. The FX variants used were non-Gla-non-EGF1 FX variants with alanine at one site (except at position 118, which is alanine in the wild type, where an alanine substitution was introduced on the run), corresponding to residues 86-448 of SEQ ID NO: 2 s An N-terminal His tag (NNNNNNH, for affinity purification) attached via a short GS linker (GGGGSGGGGS). Variants covering epitope residues as defined in Example 13 are listed in Table 9.

¹⁾ According to SEQ ID NO: 2.

²⁾ EGF2 and PD belong to the second epidermal growth factor-like domain and protease domain, respectively.

Wild-type Gla-free EGF1 FX and its variants listed in Table 9 were expressed in the HEK293 system and purified by affinity chromatography. No expression or poor purity was observed for variants L117A, L303A, P304A, and M426A, and binding assessments for these four variants were not possible.

Identification of epitope hot spot residues was carried out using a Biacore T200 instrument at 25°C. Anti-hIgG Fc antibody from the Human Antibody Capture Kit (GE Healthcare, part number BR100839) at a concentration of 2 μg/ml was immobilized onto a CM5 Series S sensor chip (GE Healthcare, part number BR100530) using standard amine immobilization chemistry. Anti-FX antibody mAb4-6934 (a monovalent variant of mAb1-6723) was administered at a flow rate of 5 μL/min. for 30 s and captured with immobilized anti-hIgG Fc antibody. Subsequently, 5 µM (with 2- or 3-fold serial dilutions) of variants T116A, A118S, T127A, F229A and E226A, 10 µM (with 5-fold serial dilutions) of variant Y230A and 10 µM (with 2- or 3-fold serial dilutions) ) WT and variants H101A, E103A, R113A, S227A, E228A, R287A, E305A, L419A, K420A, D423A, R424A, K427A and T428A were administered at a flow rate of 5 μl/min. for 90 s, allowing binding of the captured anti-FX antibody, followed by a 90 s injection of buffer to allow dissociation of non-GLA-non-EGF1 FX variants. The running buffer (also used to dilute the anti-FX antibody and the non-GLA-non-EGF1 hFX variants) contained 10 mM HEPES, 150 mM NaCl, 1 mg/ml BSA, and 5 mM CaCl ₂ (pH 7.4). The sensor chip was regenerated using 1 M formic acid. Binding data were analyzed using a steady state fit according to a 1:1 model in Biacore Evaluation 2.0 software supplied by GE Healthcare. Binding data are presented as % binding of FX variants to anti-FX antibody (monovalent mAb1-6723) relative to binding of wild-type FX to anti-FX antibody when administered at 5 or 10 μM of FX variants and are calculated according to the formula:

Binding (%)=100% × [(R _{max_FXvar,Ab} )/(R _{max_Ab} )] / [(R _{max_FXwt,Ab} )/(R _{max_Ab} )],

where R _{max_Ab} represents the capture level (RU) of the anti-FX antibody, and R _{max_FXvar,Ab} and R _{max_FXwt,Ab} represent the binding (RU) of the FX and wild-type FX variants at the same concentration (5 μM for all except the Y230A variant, whose concentration was 10 μM) with captured anti-FX antibody, respectively. The results are shown in Table 10.

Hot spot remnants for mAb1-6723

Hot spot residues for mAb1-6723 are defined as positions at which substitution of a wild-type residue with alanine (or substitution at position 118 of alanine with serine) reduces antibody binding to 30% or less relative to antibody binding to wild-type FX at 5 μM WT ( or variant) FX without GLA-without EGF1.

Hotspot residues for mAb1-6723 (experimentally represented by its monovalent analogue, mAb4-6934):

R113, Y230, K420, D423, R424 and K427.

Example 15: Identification of Hot Spot Remnants on FIX/FIXa

To identify residues critical for the interaction (referred to as the “hot spot”) between the FIX/FIXa Abs mAb0-1886, mAb0-1998 and mAb1-1307 and FIX, a set of FIX variants was selected based on the crystal structure of FIXa in complex with the corresponding Fab fragments (Fab7237, Fab7238 and Fab7236, respectively). As detailed below, selected FIX variants were transiently expressed in mammalian cells, purified, and characterized for their binding to the monovalent variants mAb0-1886, mAb0-1998, and mAb1-1307 using surface plasmon resonance (SPR).

Generation of FIX mutants

A DNA plasmid suitable for transient expression in mammals was constructed using an expression cassette encoding amino acid residues 1-461 of human FIX (UNIPROT P00740, excluding the T194A mutation by UNIPROT numbering, corresponding to T148A of SEQ ID NO: 1), followed immediately by followed by six histidine residues (6×His tag, for affinity purification). The secreted mature FIX protein chain produced using this construct is identical to the A148 allelic form of human FIX (Anson et al. EMBO J. 1984 3:1053-1060, McGrawetal, Proc Natl Acad Sci USA. 1985 82:2847-2851) except adding a C-terminal His tag.

Using the construct as a template, selected mutations were introduced by PCR. For each point mutation listed in Table 11, a forward primer containing the required amino acid substitution and a reverse primer without amino acid mutations were created. These primers were used in a standard PCR reaction with the vector described above as a template to amplify the complete vector sequence. Ligation-free cloning was used to join the ends of the resulting amplified DNA fragment into a circular expression plasmid using overlapping sequences introduced by forward and reverse primers.

The looped plasmids were transformed into E. coli cells, grown on selective agar plates to form colonies, and the colonies were used to start liquid E. coli cultures. After E. coli cultures grew overnight, plasmid preparations were prepared and mutants were identified by DNA sequencing.

Recombinant protein production was achieved by transfecting expi293F cells growing in suspension culture in Expi293 Expression™ medium (ThermoFisher Scientific, catalog number A1435101), using the ExpiFectamine™ 293 transfection kit (ThermoFisher Scientific, catalog number A14525) and plasmid DNA encoding each of the required FIX variants, as well as wild-type FIX (corresponding to SEQ ID NO: 1 with a C-terminal His tag). Vitamin K was added to a final concentration of 5 mg/ml at the time of transfection. Transfection enhancers 1 and 2 from the ExpiFectamine™ 293 transfection kit were added the day after transfection. Cell cultures were harvested 5 days after transfection by centrifugation.

The C-terminal His tag in each FIX variant was used to purify the protein in a multiwell robotic batch unit. Briefly, collected cell culture supernatants were brought to binding conditions, mixed with Ni Sepharose 6 Fast Flow affinity purification resin (GE Healthcare, part number 17-5318-02, 50 μl resin pelleted/ml cell culture medium) and incubated with shaking within 20 minutes. The resin/supernatant mixtures were then transferred to a filter plate and the liquid was passed through the filter plate using a vacuum. The resin remaining on the filter plate was washed three times before eluting in high imidazole buffer.

Determination of the concentration of purified protein solutions was performed by ELISA using anti-FIX antibody for detection and high-purity recombinant wild-type FIX for standard curves.

³⁾ According to SEQ ID NO: 1.

⁴⁾ EGF2 and PD belong to the second epidermal growth factor-like domain and protease domain, respectively.

*Variants marked with an asterisk show very low levels of expression and cannot be assessed in binding studies.

Thermal stability of FIX options

To test whether the introduction of amino acid substitutions in FIX variants leads to destabilization and misfolding, the midpoint ( _Tm ) of the thermal unfolding conformational transition was determined for the variants.

Purified FIX variants were loaded into standard capillaries (Prometheus NT.48 nanoDSF Grade standard capillaries, Nanotemper Technologies GmbH, Munich) and injected into Prometheus NT.48 (Nanotemper Technologies GmbH, Munich). The excitation intensity was 70%, and thermal unfolding followed at 20-90°C with a heating rate of 1.5°C/min. Tryptophan fluorescence was measured by excitation at 280 nm and recording emission at 330 nm and 350 nm. It is possible to determine the T _m of FIX variants (except when the protein concentration was below 20 μg/ml, which was the case for FIX N101D, H256A, L330A, S339A, G393I, Y404A and N406Q) by the ratio of fluorescence measured at 350 nm and 330 nm (F350/F330). In all cases, the PR.ThermControl v2.0.4 program (NanoTemper Technologies GmbH, Munich) was able to automatically select T _m by determining the maximum of the first derivative of the F350/F330 unfolding curve. The _Tm for wild type FIX was found to be 51°C, and the _Tm for variants was found to range from 47 to 54°C, indicating that amino acid substitutions do not cause significant destabilization.

SPR Analysis

FIX variants were characterized for their binding to mAb0-1886, mAb0-1998 and mAb1-1307 using surface plasmon resonance (SPR) by capturing the FIX variant via a C-terminal His tag. To avoid potential avidity effects associated with traditional bivalent antibody, i.e. to ensure a 1:1 interaction, monovalent variants of mAb0-1886, mAb0-1998 and mAb1-1307, designated mAb4-0673, mAb4-0004 and mAb3-3279, respectively (prepared as described in Example 7), were used as analytes.

SPR analyzes were performed on Biacore 4000 or Biacore T200 instruments (Biacore AB, Uppsala, Sweden). For experiments on the T200 device, the following conditions were used: measurements were carried out at a temperature of 25°C. Anti-His antibody at a concentration of 25 μg/ml (R&D Systems, catalog number MAB050) was immobilized on the CM5 sensor chip using standard amine immobilization chemistry. Anti-FIX antibody variants at 25 nM were injected at a flow rate of 10 μL/min. within 1 min. and captured via their His tag with an immobilized anti-His antibody. Subsequently, 200 nM (with 4-fold serial dilutions), 1600 nM (with 3-fold serial dilutions), and 2000 nM (with 3-fold serial dilutions) mAb4-0004, mAb3-3279, and mAb4-0673, respectively, were administered at a flow rate of 50 µl/min. for 5 minutes, allowing binding of the captured FIX variant, followed by a 10-minute injection of buffer to allow monovalent anti-FIX antibodies to dissociate. The working buffer was 20 mM Tris, 150 mM NaCl, 5 mM _CaCl2 , 0.05% Tween-20, 1 mg/ml BSA, pH 7.4. It was also used to dilute FIX antibody samples and FIX samples. Chip regeneration was carried out using 10 mM glycine, pH 2.0. Binding data were analyzed according to a 1:1 model using BiaEvaluation 4.1 provided by the manufacturer (Biacore AB, Uppsala, Sweden). A similar experimental setup was used for the Biacore 4000 instrument.

Initially, all FIX variants listed in Table 11 were screened using the Biacore 4000 instrument for binding to all three monovalent antibodies, mAb4-0004, mAb3-3279 and mAb4-0673. Antibody binding to FIX variants containing mutations at positions corresponding to their respective epitope residues (defined by the distance criterion as described in Example 6) was, as expected, impaired to varying degrees. No significant effect on antibody binding was observed for FIX variants containing mutations at positions inconsistent with their corresponding epitope residues. In particular, none of the substitutions made in the EGF2 domain had any effect on binding to any antibody (data not shown).

More detailed binding analysis was performed on residues identified as epitope residues (see Example 6) using a Biacore T200 instrument. The results are shown in Table 12.

Binding data are presented as % antibody binding to the FIX variant relative to antibody binding to wild-type FIX, calculated using the formula:

Binding (%) = 100% × [(R _{max_Ab,FIX_var} )/(R _{max_FIXvar} )] / [(R _{max_Ab,FIX_wt} )/(R _{max_FIXwt} )],

where R _{max_FIXvar} and R _{max_FIXwt} represent the capture rate (RU) of the FIX variant and wild-type FIX, respectively, and where R _{max_Ab,FIX_var} and R _{max_Ab,FIX_wt} represent the binding (RU) of the antibody to the captured FIX variant and wild-type FIX, respectively. The results are shown in Table 12.

1) Position according to SEQ ID NO: 1.

2) 100% × [(R _{max_Ab,FIX_var} ) / (R _{max_FIXvar} )] / [(R _{max_Ab,FIX_wt} ) / (R _{max_FIXwt} )].

Hot spot residues for mAb1-1307, mAb0-1998 and mAb0-1886

Hot spot residues for mAb1-1307, mAb0-1998, and mAb0-1886 are defined as positions at which replacement of the wild-type residue with alanine reduces antibody binding to 30% or less relative to antibody binding to wild-type FIX.

Hot spot residues for mAb1-1307 (experimentally represented as mAb3-3279):

H257, K293 and N406.

Hot spot residues for mAb0-1998 (experimentally represented as mAb4-0004):

R338 and K341.

Hot spot residues for mAb0-1886 (experimentally represented as mAb4-0673):

D332, R333, L337 and R338.

For both mAb0-1998 and mAb0-1886, the residue most conducive to binding is R338; substitution of R338 with alanine (R338A) in FIX showed the greatest effect on antibody binding, which was significantly reduced to 2% and 3% for mAb0-1998 and mAb0-1886, respectively, relative to antibody binding to wild-type FIX.

Example 16. Activity of bispecific antibodies to FIX(a)/FX(a) in the FXa production assay

The procoagulant activity of FIXa/FX bispecific antibodies was determined based on their ability to stimulate FX activation by FIXa in the presence of a procoagulant phospholipid membrane. The bispecific antibodies (BiAbs) tested are listed in Table 13 and ACE910 was included for comparison.

The procoagulant activity of each bispecific antibody is described as the fold stimulation compared to FX activation by free FIXa at a given antibody concentration. Bispecific antibodies were tested at 8 concentrations (prepared by serial threefold dilutions in assay buffer) by pre-incubation with 125 pM FIXa derived from human plasma (Haematologic Technologies Inc., USA) and 500 µM phospholipid vesicles of a 25:75 phosphatidylserine:phosphatidylcholine mixture (Haematologic Technologies Inc, USA) in assay buffer (50 mM HEPES, 100 mM NaCl, 5 mM CaCl2, _0.1 % (w/v) PEG8000, pH 7.3, +1 mg/ml BSA) for 10 min. Activation was then initiated by adding FX derived from human plasma (Haematologic Technologies Inc, USA) to a concentration of 25 nM. After 15 min activation at room temperature, the reaction (50 μl) was quenched by adding 25 μl quenching buffer (50 mM HEPES, 100 mM NaCl, 60 mM EDTA, 0.1% PEG8000, pH 7.3, + 1 mg/ml BSA ). The amount of FXa produced was determined by adding 25 μl of 2 mM chromogenic substrate S-2765 (Chromogenix, Sweden) and measuring the conversion of the chromogenic substrate by measuring the optical density at 405 nm (ΔOD/min) in a microplate reader. Similarly, FX activation by free FIXa was determined at a FIXa concentration of 25 nM and a reaction time of 60 min.

The measured activity was normalized according to the concentration of FIXa present in the assay and the reaction time. By dividing this number by the similarly normalized rate of FXa production in the absence of antibody, the fold stimulation by antibody at a given concentration was calculated.

Thus, the biAb stimulation calculation can be described as follows:

stimulation biAb = ( _AFIXa+biAb / ([FIXa] _analysis × t _reaction )) / A _FIXa,norm. ,

where A _FIXa+biAb represents the activity measured in the presence of the bispecific antibody, [FIXa] _assay represents the concentration of FIXa in the assay, t _reaction represents the reaction time, and A _FIXa,norm. represents normalized free FIXa activity.

Table 13 lists the maximum stimulation determined for each bispecific antibody among the 8 antibody concentrations tested, as well as the concentration at which maximum stimulation was observed. It was found that for all bispecific antibodies tested, the maximum stimulation was higher than that measured for ACE910, which was tested in the concentration range from 0 to 15300 nM.

Example 17. Activity of bispecific antibodies to FIX(a)/FX(a) in the thrombin generation test (TGT) in platelet-poor or platelet-rich plasma of a person with simulated hemophilia A

The procoagulant activity of the bispecific antibodies mAb4-7761, mAb4-7762, mAb4-7789, mAb5-0057 and mAb5-1409 (see Table 14) was determined based on their ability to stimulate thrombin formation in the presence of either a procoagulant synthetic phospholipid membrane or platelets in accordance with the principles , described by Hemker et al. (Pathophysiol Haemost Thromb 2002;32:249-253). ACE910 was included for comparison. Each antibody (test compound) was tested in a thrombin generation test (TGT) in pooled blood plasma from platelet-poor hemophilia A (HA) patients (HA-PPP) and/or human platelet-rich plasma with induced HA (HA- PRP).

Hemophilia A-induced platelet-rich plasma (HA-PRP)

Blood was obtained from healthy, consenting donors by venipuncture. Six volumes of blood were collected with 1 volume of acid citrate dextrose (ACD; 85 mM sodium citrate, 110 mM dextrose, and 62.3 mM citric acid, pH 4.9), final pH 6.5, and centrifuged for 20 min. at 220 g at room temperature (RT). Platelet-rich plasma (PRP) was collected and platelet concentrations were determined using a Medonic CA 620 hematology analyzer (Boule Diagnostics AB, Sponga, Sweden). The part of the blood plasma containing erythrocytes was centrifuged for another 10 minutes. at 600 g at RT. Platelet-poor plasma (PPP) was collected and used to dilute PRP to 300,000 platelets/μL. NA conditions were induced by adding anti-human FVIII-neutralizing antibody (sheep anti-human factor VIII antibody - 5 mg, Haematologic Technologies, Vermont, USA) to a final concentration of 0.1 mg/ml and stirred gently at 2 rpm. for 30 minutes at RT.

Thrombin formation test

Thrombin generation tests (TGT) in HA-PRP and HA-PPP (George King Bio-Medical Inc, Kansas, USA) were performed using a standard-calibrated automated thrombography using a 96-well plate fluorimeter (Fluoroscan Ascent FL, Thermolabsystems, Helsinki, Finland ). Reaction mixtures contained 70 μl HA-PRP (300,000 platelets/μl) or HA-PPP, 10 μl test compound dilution (diluted in 20 mM HEPES, 140 mM NaCl, pH 7.4, 2% BSA), 20 μl CAT reagents, containing tissue factor (TF) (PRP reagent; TF without synthetic phospholipids, PPP LOW reagent; TF with synthetic phospholipids, 1 pM final TF, Thrombinoscope BV, Maastricht, the Netherlands) or thrombin calibrator (Thrombinoscope BV), and 20 μl of a mixture containing fluorescently labeled thrombin substrate z-Gly-Gly-Arg-AMC (3 mM) and CaCl ₂ . (90 mM) (Thrombinoscope BV). TGT was performed at eight concentrations of test compound (0.3, 1.0, 3, 10, 30, 100, 300 and 900 nM, final plasma concentration) or with the addition of buffer alone (20 mM HEPES, 140 mM NaCl, pH 7.4, 2% BSA) (represents AN control). Concentration ranges were tested in at least three independent experiments in HA-PPP from the same stock or in blood from four different donors. Normal control levels in TGT were measured using untreated human PRP or CRYOcheck™ pooled normal human PRP blood plasma (Precision Biologic Inc., Dartmouth, Canada) supplemented with buffer only (20 mM HEPES, 140 mM NaCl, pH 7.4, 2% BSA). In total, TGT was performed for 90 minutes and the TGT parameter of peak thrombin height (nM) was analyzed using Thrombinoscope software (Thrombinoscope BV).

Figure 2/Table 15 shows the measured peak thrombin generation rates for each bispecific antibody at the concentrations tested in HA-PPP. The data show that all tested compounds increase peak thrombin generation above the level observed in the absence of antibody, i.e. exhibit procoagulant activity. In addition, thrombin generation levels of 30 to 300 nM for mAb4-7761, mAb4-7762, mAb4-7789, mAb5-0057, and mAb5-1409 are higher than those observed for ACE910, demonstrating superior potency. In addition, the thrombin generation levels at 300-900 nM mAb5-0057 and mAb5-1409 are higher than those observed at 900 nM ACE910, demonstrating higher potency and efficacy values for these compounds compared to ACE910.

Figure 3/Table 16 shows the measured peak thrombin generation levels for mAb5-0057 and mAb5-1409 at the concentrations tested in HA-PRP. Under these conditions, mAb5-0057 and mAb5-1409 also demonstrated superior potency and efficacy values compared to ACE910.

Example 18. Activity of monovalent partial (OA) antibodies to FIX/FIXa in the thrombin generation test (TGT) in platelet-poor human plasma in hemophilia A

The procoagulant activity of the monovalent partial (OA) variant of mAb1-9016 was determined based on its ability to stimulate thrombin formation in the presence of a procoagulant phospholipid membrane according to the principles described by Hemker et al. (2002) Pathophysiol Haemost Thromb, 32:249–253. A partial version of antibody 224F3 (mAb1-1582) was included for comparison. Each antibody (test compound) was tested in a thrombin generation test (TGT) in pooled blood plasma from hemophilia A (HA) platelet-poor (HA-PPP) patients (George King Bio-Medical Inc., Kansas, USA) using a calibrated standard automated thrombography using a 96-well plate fluorimeter (Fluoroscan Ascent FL, Thermolabsystems, Helsinki, Finland). The reaction mixtures contained 70 μl HA-PPP, 10 μl test compound (diluted in 20 mM HEPES, 140 mM NaCl, pH 7.4, 2% BSA), 20 μl PRP reagents (synthetic phospholipids, Thrombinoscope BV, Maastricht, the Netherlands), containing activated human plasma-derived factor XI (hFXIa, 8.3 mU/ml total) (Enzyme Research Laboratories, Indiana, USA) or thrombin calibrator (Thrombinoscope BV), and 20 μl of a mixture containing fluorescently labeled thrombin substrate Z-Gly-Gly-Arg-AMC (3 mM) and CaCl ₂ (90 mM) (Thrombinoscope BV). TGT was performed at five concentrations of test compound (30, 100, 300, 600 and 900 nM, final plasma concentration) or with the addition of buffer alone (20 mM HEPES, 140 mM NaCl, pH 7.4, 2% BSA) (representing NA control). The concentration range was tested in two independent experiments in HA-PPR from the same stock. In total, TGT was performed for 90 minutes and the TGT parameter of peak thrombin height (nM) was analyzed using Thrombinoscope software (Thrombinoscope BV). Figure 4 and Table 17 show/list the measured peak thrombin generation rates for each monovalent partial antibody at the concentrations tested. Data show that the OA variant of mAb1-9016 increases peak thrombin generation above the level observed in the absence of antibody, i.e. exhibits procoagulant activity. In addition, thrombin formation induced by the OA variant of antibody mAb1-9016 is higher than that observed for the monovalent OA variant of antibody 224F3 (mAb1-1582).

Example 19 Binding Affinities Determined by Isothermal Titration Calorimetry (ITC)

Binding affinities for anti-FIX/FIXa antibodies and anti-FX/FXa antibodies binding to FIX/FIXa and FX/FXa, respectively, were measured by isothermal titration calorimetry (ITC) using a PEAQ-ITC calorimeter (Malvern, UK). Experiments were carried out at 37°C and pH 7.4 using 25 mM Tris, 150 mM NaCl, 5 mM CaCl ₂ (Tris buffer). The sample well (200 μl) contains either FIX, FIXa, FX or FXa, and anti-FIX/FIXa and anti-FX/FXa antibodies are injected via a syringe. All proteins are dialyzed extensively in Tris buffer before measurements to ensure suitable buffer conditions. The thermal equilibration step was followed by a 60-second delay and then an initial injection of 0.2 μl of antibody followed by 14 injections of 2.5 μl of antibody at 120-s intervals. The stirring speed is maintained at 750 rpm, and the reference power is kept constant at 5-10 μcal/s. The heat associated with each antibody injection is integrated and plotted against the molar ratio of ligand to macromolecule. The resulting isotherm is fitted to a single site binding model to obtain affinity (KD), stoichiometry (n) and enthalpy of interaction (ΔH) using software provided by the manufacturer. Experiments were carried out in two or three repetitions.

While certain features of the present invention have been illustrated and described herein, many modifications, substitutions, alterations and equivalents will be apparent to those skilled in the art. Therefore, it is to be understood that the appended claims are intended to cover all such modifications and changes that are in the true spirit of the present invention.

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LIST OF SEQUENCES

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65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 6

<211> 107

<212> Protein

<213> Homo sapiens

<400> 6

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 7

<211> 120

<212> Protein

<213> Homo sapiens

<400> 7

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Val Ser Gly Ser Ile Gly Tyr Ala Arg Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Gly Gln Tyr Asp Glu Asp Ala Gly Met Asp Val Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 8

<211> 106

<212> Protein

<213> Homo sapiens

<400> 8

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Arg Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Tyr Pro Thr Trp Thr

85 90 95

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 9

<211> 120

<212> Protein

<213> Homo sapiens

<400> 9

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Lys Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Gly Gln Tyr Asp Glu Asp Ala Gly Met Asp Val Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 10

<211> 106

<212> Protein

<213> Homo sapiens

<400> 10

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Trp Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Pro Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Tyr Pro Thr Trp Thr

85 90 95

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 11

<211> 119

<212> Protein

<213> Homo sapiens

<400> 11

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Glu Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 12

<211> 112

<212> Protein

<213> Homo sapiens

<400> 12

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Arg Leu Leu His Ser

20 25 30

Thr Gly Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Arg Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 13

<211> 119

<212> Protein

<213> Homo sapiens

<400> 13

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Arg Phe Thr Ser Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Thr Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Met Ser Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 14

<211> 112

<212> Protein

<213> Homo sapiens

<400> 14

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 15

<211> 122

<212> Protein

<213> Homo sapiens

<400> 15

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Gly Asp Asn Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 16

<211> 107

<212> Protein

<213> Homo sapiens

<400> 16

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 17

<211> 120

<212> Protein

<213> Homo sapiens

<400> 17

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Gly Gln Tyr Asp Glu Asp Ala Gly Met Asp Val Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 18

<211> 106

<212> Protein

<213> Homo sapiens

<400> 18

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Tyr Pro Thr Trp Thr

85 90 95

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 19

<211> 119

<212> Protein

<213> Homo sapiens

<400> 19

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Met Glu Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 20

<211> 112

<212> Protein

<213> Homo sapiens

<400> 20

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Gly Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 21

<211> 120

<212> Protein

<213> Homo sapiens

<400> 21

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Thr Ser

20 25 30

Trp Ile Val Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Phe Thr Ser Tyr Ser Pro Ser Phe

50 55 60

Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Leu His Tyr Tyr His Ser Glu Glu Phe Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 22

<211> 109

<212> Protein

<213> Homo sapiens

<400> 22

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Arg Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Phe Gly Ser Ser Arg

85 90 95

Leu Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 23

<211> 120

<212> Protein

<213> Homo sapiens

<400> 23

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Ala Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Thr Leu Tyr Thr Gly Gly Ser Thr His Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Ala Thr Trp Glu Leu Leu Ser Ile Val Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 24

<211> 112

<212> Protein

<213> Homo sapiens

<400> 24

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Asp Gly Tyr Thr Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Thr Pro Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 25

<211> 121

<212> Protein

<213> Homo sapiens

<400> 25

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Asn

20 25 30

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Thr Ile Gly Gly Ser Gly Gly Arg Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Ser Trp Gly Phe Gly Trp Tyr Phe Asp Leu Trp Gly

100 105 110

Arg Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 26

<211> 109

<212> Protein

<213> Homo sapiens

<400> 26

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Gly Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Asp Thr Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gln Tyr Gly Thr Ser

85 90 95

Pro Gly Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 27

<211> 122

<212> Protein

<213> Homo sapiens

<400> 27

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Ala Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Pro His Asn Gly Asn Thr His Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Ser Leu Trp Phe Gly Glu Leu Leu Pro Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 28

<211> 108

<212> Protein

<213> Homo sapiens

<400> 28

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Val Ser Ser Ser

20 25 30

Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Ala Val Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Tyr Gly Ser Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 29

<211> 120

<212> Protein

<213> Homo sapiens

<400> 29

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Lys Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Thr Asp Ser Tyr Thr Thr Tyr Ser Pro Ser Leu

50 55 60

His Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Gln Leu Trp Leu Arg Asp Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 30

<211> 107

<212> Protein

<213> Homo sapiens

<400> 30

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Val Gly Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Arg Ala Ala Gly Ile Pro Asp Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Tyr Trp Pro Ala

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 31

<211> 121

<212> Protein

<213> Homo sapiens

<400> 31

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr

20 25 30

Trp Ile Val Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Ser Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Ser Gln Arg Arg Gly Tyr Leu Asp Ala Phe Asp Ile Trp Gly

100 105 110

Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 32

<211> 108

<212> Protein

<213> Homo sapiens

<400> 32

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Gly Ile Gly Thr Asp

20 25 30

Leu Ala Trp Tyr Gln Gln Asn Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ala Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Leu

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 33

<211> 118

<212> Protein

<213> Homo sapiens

<400> 33

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Thr Tyr Ser Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Pro Ser Phe Thr Asp His Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 34

<211> 106

<212> Protein

<213> Homo sapiens

<400> 34

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Ala Ser Ala Arg Ala Ala Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu

65 70 75 80

Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Asn Trp Pro Ala Thr

85 90 95

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 35

<211> 121

<212> Protein

<213> Homo sapiens

<400> 35

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Asp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala His Ser Glu Asp Tyr Tyr Asp Ser Ser Gly Phe Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 36

<211> 108

<212> Protein

<213> Homo sapiens

<400> 36

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ala Arg Ala Ala Gly Ile Pro Asp Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Asn Trp Pro Pro

85 90 95

Val Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 37

<211> 126

<212> Protein

<213> Homo sapiens

<400> 37

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr

20 25 30

Ala Leu Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Asp Trp Val

35 40 45

Ser Thr Ile Ser Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Ser

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Asp Glu Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gly Gly Phe Ala Ser Ser Ser Gly Arg Tyr Tyr Tyr Tyr Ala

100 105 110

Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120 125

<210> 38

<211> 107

<212> Protein

<213> Homo sapiens

<400> 38

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp

20 25 30

Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 39

<211> 121

<212> Protein

<213> Homo sapiens

<400> 39

Gln Ala Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Thr Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Glu Gln Trp Leu Val Pro Asp Asp Ala Phe Asp Ile Trp Gly

100 105 110

Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 40

<211> 113

<212> Protein

<213> Homo sapiens

<400> 40

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Ser Asn Lys Asn Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Asn Leu Ile Ile Tyr Trp Ala Ser Asn Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys His Gln

85 90 95

Tyr Tyr Asn Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 41

<211> 123

<212> Protein

<213> Homo sapiens

<400> 41

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Leu Arg Ser Tyr

20 25 30

Gly Ile Ser Trp Leu Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Asp Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Leu Thr Thr Glu Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Met Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Glu Asp Ile Ser Ser Arg Trp Ser Pro Asp Thr Phe Asp Ile

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 42

<211> 107

<212> Protein

<213> Homo sapiens

<400> 42

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Tyr Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn His Tyr Ser Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 43

<211> 124

<212> Protein

<213> Homo sapiens

<400> 43

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Asn

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Asp Trp Ala Met Val Arg Gly Val Ile Thr Asn Ala Phe Asp

100 105 110

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 44

<211> 106

<212> Protein

<213> Homo sapiens

<400> 44

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Arg Tyr Ser Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 45

<211> 120

<212> Protein

<213> Homo sapiens

<400> 45

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Asn

20 25 30

Tyr Tyr Tyr Trp Gly Trp Ile Arg Gln Thr Pro Gly Lys Arg Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Ile Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Thr Leu Ser Phe Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Glu Ala Glu Arg His Asp Ser Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 46

<211> 107

<212> Protein

<213> Homo sapiens

<400> 46

Glu Ile Val Met Thr Gln Ser Pro Val Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Val Tyr Asn Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Phe Pro Gly Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Val Trp Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 47

<211> 120

<212> Protein

<213> Homo sapiens

<400> 47

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Tyr Tyr Tyr Trp Gly Trp Ile Arg Gln Thr Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Ile Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Thr Leu Ser Phe Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Glu Ala Glu Arg His Asp Ser Phe Asp Leu Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 48

<211> 107

<212> Protein

<213> Homo sapiens

<400> 48

Glu Ile Val Met Thr Gln Ser Pro Val Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Val Tyr Asn Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Gly Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Val Trp Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 49

<211> 116

<212> Protein

<213> Homo sapiens

<400> 49

Gln Glu Gln Leu Val Glu Ser Gly Gly Asp Val Val Gln Pro Gly Lys

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Thr Ser Gly Phe Thr Phe Ser Phe Tyr

20 25 30

Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Ser Ile Ser Tyr Asp Gly Thr Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Gly Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Lys Asp Ser Arg Gly Met Ala Val Trp Gly Gln Gly Thr Thr Val

100 105 110

Thr Val Ser Ser

115

<210> 50

<211> 112

<212> Protein

<213> Homo sapiens

<400> 50

Asp Val Leu Met Thr Gln Ser Pro Leu Phe Leu Pro Val Thr Phe Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Arg Ser Leu Val Tyr Ser

20 25 30

Asp Gly Asn Thr Tyr Val Asn Trp Phe His Gln Arg Pro Gly Gln Ser

35 40 45

Pro Arg Arg Leu Ile Tyr Lys Val Ser Asn Arg Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly

85 90 95

Thr His Trp Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 51

<211> 120

<212> Protein

<213> Homo sapiens

<400> 51

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Tyr Tyr Tyr Trp Gly Trp Ile Arg Gln Thr Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn His Phe

65 70 75 80

Ser Leu Asn Leu Ser Phe Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Glu Ala Glu Arg His Asp Thr Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 52

<211> 107

<212> Protein

<213> Homo sapiens

<400> 52

Glu Ile Val Met Thr Gln Ser Pro Val Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Ile Ser Asn Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Gly Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 53

<211> 119

<212> Protein

<213> Homo sapiens

<400> 53

Gln Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser

20 25 30

Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu

35 40 45

Trp Leu Ala Leu Ile Tyr Trp Asp Asp Asp Lys Arg Tyr Ser Pro Ser

50 55 60

Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val

65 70 75 80

Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr

85 90 95

Cys Ala His Arg Leu Gly Ser Tyr Glu Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 54

<211> 112

<212> Protein

<213> Homo sapiens

<400> 54

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Asn Gly Tyr Ser Tyr Leu Asp Trp Tyr Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala

85 90 95

Leu Gln Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 55

<211> 117

<212> Protein

<213> Homo sapiens

<400> 55

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Pro Asn Ser Gly Ser Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Trp Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Arg Leu Arg Ser Asp Ala Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Tyr Tyr Asp Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 56

<211> 107

<212> Protein

<213> Homo sapiens

<400> 56

Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys

1 5 10 15

Glu Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Ile Gly Asn Ser

20 25 30

Leu His Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Leu Leu Ile

35 40 45

Glu Tyr Ala Ser Gln Ser Ile Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ile Ser Leu Glu Ala

65 70 75 80

Glu Asp Ala Ala Ala Tyr Tyr Cys His Gln Ser Ser Arg Leu Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 57

<211> 124

<212> Protein

<213> Homo sapiens

<400> 57

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Lys Ile Ser Cys Lys Gly Phe Gly Tyr Arg Phe Thr Asp Tyr

20 25 30

Trp Ile Gly Trp Ile Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Tyr Pro Asp Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe

50 55 60

Gln Gly Gln Val Thr Ile Ser Ala Asp Arg Ser Val Lys Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg His Gly Ile Ile Gly Thr Arg Asp Tyr Tyr Tyr Gly Met Asp

100 105 110

Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 58

<211> 107

<212> Protein

<213> Homo sapiens

<400> 58

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Asn Asn Tyr

20 25 30

Leu Asp Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Thr Leu Ile Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Tyr Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Ile Ala Thr Tyr Tyr Cys Gln His Tyr Asp Asn Leu Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 59

<211> 124

<212> Protein

<213> Homo sapiens

<400> 59

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Gly Gly Ser Lys Glu Tyr Ala Ala Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Phe Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Asp Trp Ala Met Val Arg Gly Val Ile Thr Asn Val Phe Asp

100 105 110

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 60

<211> 106

<212> Protein

<213> Homo sapiens

<400> 60

Ala Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Ser Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 61

<211> 119

<212> Protein

<213> Homo sapiens

<400> 61

Gln Glu Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Phe Asn Thr Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Val Ile Ser Phe Asp Ser Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Glu Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Val Lys Gly Arg Ser Tyr Tyr Trp Tyr Phe Asp Leu Trp Gly Arg Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 62

<211> 112

<212> Protein

<213> Homo sapiens

<400> 62

Asn Ile Ile Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asp Gly Asn Thr Tyr Leu Thr Trp Leu Gln Gln Arg Pro Gly Gln Pro

35 40 45

Pro Arg Leu Leu Ile Tyr Lys Ile Ser Lys Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr Cys Met Gln Ala

85 90 95

Thr His Phe Pro His Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 63

<211> 121

<212> Protein

<213> Homo sapiens

<400> 63

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Tyr Trp Val

35 40 45

Ser Ala Ile Ser Gly Asn Gly Asp Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Glu Glu Gly Ile Val Val Ala His Asp Ala Phe Glu Ile Trp Gly

100 105 110

Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 64

<211> 113

<212> Protein

<213> Homo sapiens

<400> 64

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Ile Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Ile Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Gly Thr Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile

100 105 110

Lys

<210> 65

<211> 109

<212> Protein

<213> Oryctolagus cuniculus

<400> 65

Gln Glu Arg Leu Lys Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr

1 5 10 15

Pro Leu Thr Leu Thr Cys Thr Ala Gly Gly Ser Asp Ile Ser Val Tyr

20 25 30

Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Trp Ile Gly Ala Gly Gly His Ala Tyr Tyr Ala Arg Trp Ala Val

50 55 60

Gly Arg Phe Thr Ile Ser Arg Thr Ser Thr Thr Val Asp Leu Lys Met

65 70 75 80

Thr Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Ser Ala

85 90 95

Gly Leu Trp Gly Pro Gly Thr Leu Val Thr Val Ser Ser

100 105

<210> 66

<211> 112

<212> Protein

<213> Oryctolagus cuniculus

<400> 66

Gln Val Leu Thr Gln Thr Glu Ser Pro Val Ser Ala Ala Ile Gly Gly

1 5 10 15

Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Ser Val Val Asn Asp Asn

20 25 30

Arg Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Arg Leu

35 40 45

Ile Tyr Asp Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Lys

50 55 60

Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Val Gln

65 70 75 80

Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Leu Gly Gly Tyr Asp Cys Ser

85 90 95

Leu Asn Asp Cys Asn Gly Phe Gly Gly Gly Thr Glu Val Val Val Lys

100 105 110

<210> 67

<211> 119

<212> Protein

<213> Oryctolagus cuniculus

<400> 67

Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Thr Tyr Thr

20 25 30

Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly

35 40 45

Ile Ile Ala Ser Ser Gly His Ala Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Ala Val Asp Leu Lys Met

65 70 75 80

Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Ala

85 90 95

Leu Tyr Ser Gly Gly Gly Tyr Trp Pro Gly Gly Phe Trp Gly Pro Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 68

<211> 111

<212> Protein

<213> Oryctolagus cuniculus

<400> 68

Ile Val Met Thr Gln Thr Pro Ser Ser Lys Ser Val Pro Val Gly Asp

1 5 10 15

Thr Val Thr Ile Asn Cys Gln Ala Ser Lys Ser Val Tyr Ser Asn Asn

20 25 30

Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Arg

50 55 60

Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Val Val

65 70 75 80

Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Tyr Glu Glu Ser Ile

85 90 95

Asp Asp Asp Thr Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys

100 105 110

<210> 69

<211> 122

<212> Protein

<213> Homo sapiens

<400> 69

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Thr Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Gly Asp Asn Ile Gly Tyr Val Glu Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Asp Lys Asn Ser Leu His

65 70 75 80

Leu Glu Leu Asn Ser Leu Arg Pro Glu Asp Thr Ala Leu Tyr Phe Cys

85 90 95

Ala Lys Ser Tyr Gly Ser Gly Ser Tyr Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 70

<211> 107

<212> Protein

<213> Homo sapiens

<400> 70

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 71

<211> 122

<212> Protein

<213> Homo sapiens

<400> 71

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Asp Asn Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Ala Phe Gly Ser Gly Ser Tyr Tyr Asn Gly Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 72

<211> 107

<212> Protein

<213> Homo sapiens

<400> 72

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Glu Tyr Asn Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 73

<211> 122

<212> Protein

<213> Homo sapiens

<400> 73

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Asp Asn Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ile Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Ala Phe Gly Ser Gly Ser Tyr Tyr Asn Gly Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Ala Leu Val Thr Val Ser Ser

115 120

<210> 74

<211> 107

<212> Protein

<213> Homo sapiens

<400> 74

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Glu Tyr Asn Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 75

<211> 121

<212> Protein

<213> Mus musculus

<400> 75

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Ala Tyr Glu Phe Ser Lys His

20 25 30

Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val

35 40 45

Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Arg Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Tyr Gly Tyr Asp Glu Glu Tyr Tyr Gly Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 76

<211> 107

<212> Protein

<213> Mus musculus

<400> 76

Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Glu Ala Val Thr Ile Thr Cys Arg Val Asn Glu Asn Ile Asp Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val

35 40 45

Tyr Ala Ala Thr Leu Leu Ala Asp Gly Val Pro Pro Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Gln Phe Ser Leu Lys Ile Asn Ser Leu Gln Ser

65 70 75 80

Glu Asp Val Ala Arg Tyr Phe Cys Gln His Tyr Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 77

<211> 121

<212> Protein

<213> Mus musculus

<400> 77

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Lys His

20 25 30

Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Gln Ile Tyr Pro Gly Asp Gly Asp Ile Asn Tyr Asn Gly Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Tyr Gly Tyr Asn Glu Glu Tyr Tyr Gly Leu Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 78

<211> 107

<212> Protein

<213> Mus musculus

<400> 78

Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Glu Thr Val Thr Ile Thr Cys Arg Ile Ser Glu Asn Ile Asp Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val

35 40 45

Tyr Ala Ala Thr Leu Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Gln Phe Ser Leu Lys Ile Asn Ser Leu Gln Ser

65 70 75 80

Glu Asp Val Ala Arg Tyr Phe Cys Gln His Tyr Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 79

<211> 117

<212> Protein

<213> Oryctolagus cuniculus

<400> 79

Gln Ser Leu Ala Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Ala Pro Gly Phe Thr Ile Gly Gly Tyr His

20 25 30

Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

35 40 45

Leu Ile Asn Asn Gly Gly Ser Thr Ala Tyr Ala Asn Trp Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Leu Thr

65 70 75 80

Ser Leu Thr Ser Glu Gly Thr Ala Thr Tyr Phe Cys Ala Arg Trp Thr

85 90 95

Gly Tyr Ala Gly Asp Gly Tyr Gly Asp Ile Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 80

<211> 111

<212> Protein

<213> Oryctolagus cuniculus

<400> 80

Ala Asp Ile Val Met Thr Gln Thr Pro Ala Ser Val Ser Ala Ala Val

1 5 10 15

Gly Gly Thr Val Thr Phe Lys Cys Gln Ala Ser Glu Asn Ile Tyr Asn

20 25 30

Leu Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Ala Phe Asp Leu Ala Ser Gly Val Pro Ser Arg Phe Lys

50 55 60

Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Leu Glu

65 70 75 80

Ser Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Ser Tyr Tyr Tyr Ala Ser

85 90 95

Ser Ser Ser Tyr Pro Phe Gly Gly Gly Thr Glu Val Val Val Lys

100 105 110

<210> 81

<211> 122

<212> Protein

<213> Homo sapiens

<400> 81

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Ser Asp Asn Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ile Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Ala Phe Gly Ser Gly Ser Tyr Tyr Asn Gly Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Ala Leu Val Thr Val Ser Ser

115 120

<210> 82

<211> 107

<212> Protein

<213> Homo sapiens

<400> 82

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gly Gln Tyr Arg Ser Tyr Ser Arg

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 83

<211> 122

<212> Protein

<213> Homo sapiens

<400> 83

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Asn Gly Asp Asn Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 84

<211> 107

<212> Protein

<213> Homo sapiens

<400> 84

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Arg Ser Tyr Ser Arg

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 85

<211> 120

<212> Protein

<213> Oryctolagus cuniculus

<400> 85

Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ala Tyr Ala

20 25 30

Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

35 40 45

Ile Val Ser Ser Ser Asp Phe Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Glu Leu Lys Ile Thr

65 70 75 80

Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp Leu

85 90 95

Asn Tyr Asp Asp Tyr Glu Asn Tyr Tyr Gly Met Asp Pro Trp Gly Pro

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 86

<211> 113

<212> Protein

<213> Oryctolagus cuniculus

<400> 86

Asp Val Val Met Thr Gln Thr Pro Ser Ser Val Ser Ala Ala Val Gly

1 5 10 15

Gly Thr Val Thr Ile Lys Cys Gln Ser Ser Gln Ser Val Tyr Asn Tyr

20 25 30

Asn Leu Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Gly Ala Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly Val

65 70 75 80

Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Leu Gly Ser Phe Asp Asp

85 90 95

Asp Val Asp Tyr Glu Asn Ala Phe Gly Gly Gly Thr Glu Val Val Val

100 105 110

Lys

<210> 87

<211> 119

<212> Protein

<213> Oryctolagus cuniculus

<400> 87

Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Thr Tyr Thr

20 25 30

Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly

35 40 45

Ile Ile Ala Ser Ser Gly His Ala Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Ala Val Asp Leu Lys Met

65 70 75 80

Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Ala

85 90 95

Leu Tyr Ser Gly Gly Gly Tyr Trp Pro Gly Gly Phe Trp Gly Pro Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 88

<211> 110

<212> Protein

<213> Oryctolagus cuniculus

<400> 88

Val Met Thr Gln Thr Pro Ser Ser Lys Ser Val Pro Val Gly Asp Thr

1 5 10 15

Val Thr Ile Asn Cys Gln Ala Ser Lys Ser Val Tyr Ser Asn Asn Arg

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Arg Gly

50 55 60

Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Val Val Cys

65 70 75 80

Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Tyr Glu Glu Ser Ile Asp

85 90 95

Asp Asp Thr Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys

100 105 110

<210> 89

<211> 119

<212> Protein

<213> Homo sapiens

<400> 89

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Glu Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 90

<211> 112

<212> Protein

<213> Homo sapiens

<400> 90

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Arg Leu Leu His Ser

20 25 30

Thr Gly Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Arg Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 91

<211> 119

<212> Protein

<213> Homo sapiens

<400> 91

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 92

<211> 112

<212> Protein

<213> Homo sapiens

<400> 92

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Arg

20 25 30

Thr Tyr Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 93

<211> 123

<212> Protein

<213> Homo sapiens

<400> 93

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ala

20 25 30

Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Arg Ile Lys Arg Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala

50 55 60

Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asn Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Thr Thr Asp Gly Ile Ala Val Ala Gly Pro Leu Arg Val Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 94

<211> 113

<212> Protein

<213> Homo sapiens

<400> 94

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Val Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Ile Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 95

<211> 119

<212> Protein

<213> Homo sapiens

<400> 95

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 96

<211> 112

<212> Protein

<213> Homo sapiens

<400> 96

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Arg Leu Leu His Ser

20 25 30

Thr Gly Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 97

<211> 119

<212> Protein

<213> Homo sapiens

<400> 97

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Lys Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 98

<211> 112

<212> Protein

<213> Homo sapiens

<400> 98

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Arg

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Arg Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 99

<211> 119

<212> Protein

<213> Homo sapiens

<400> 99

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 100

<211> 112

<212> Protein

<213> Homo sapiens

<400> 100

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Arg

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Arg Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 101

<211> 119

<212> Protein

<213> Homo sapiens

<400> 101

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

His Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Met Ser Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 102

<211> 112

<212> Protein

<213> Homo sapiens

<400> 102

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 103

<211> 119

<212> Protein

<213> Homo sapiens

<400> 103

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Arg Phe Thr Ser Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Thr Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

His Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Met Arg Ile Met Ser Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 104

<211> 112

<212> Protein

<213> Homo sapiens

<400> 104

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Arg Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 105

<211> 119

<212> Protein

<213> Homo sapiens

<400> 105

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 106

<211> 112

<212> Protein

<213> Homo sapiens

<400> 106

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Arg

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Arg Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 107

<211> 119

<212> Protein

<213> Homo sapiens

<400> 107

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 108

<211> 112

<212> Protein

<213> Homo sapiens

<400> 108

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 109

<211> 119

<212> Protein

<213> Homo sapiens

<400> 109

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 110

<211> 112

<212> Protein

<213> Homo sapiens

<400> 110

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 111

<211> 119

<212> Protein

<213> Homo sapiens

<400> 111

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Arg Phe Thr Ser Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Thr Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

His Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Met Arg Ile Met Ser Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 112

<211> 112

<212> Protein

<213> Homo sapiens

<400> 112

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 113

<211> 119

<212> Protein

<213> Homo sapiens

<400> 113

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 114

<211> 112

<212> Protein

<213> Homo sapiens

<400> 114

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Arg Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 115

<211> 119

<212> Protein

<213> Homo sapiens

<400> 115

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 116

<211> 112

<212> Protein

<213> Homo sapiens

<400> 116

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Arg Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 117

<211> 119

<212> Protein

<213> Homo sapiens

<400> 117

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 118

<211> 112

<212> Protein

<213> Homo sapiens

<400> 118

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 119

<211> 119

<212> Protein

<213> Homo sapiens

<400> 119

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Arg Phe Thr Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Thr Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ser Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 120

<211> 112

<212> Protein

<213> Homo sapiens

<400> 120

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 121

<211> 122

<212> Protein

<213> Homo sapiens

<400> 121

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Arg Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Ala Gln Ile Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 122

<211> 107

<212> Protein

<213> Homo sapiens

<400> 122

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Lys Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Arg Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 123

<211> 122

<212> Protein

<213> Homo sapiens

<400> 123

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Lys Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ile Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 124

<211> 107

<212> Protein

<213> Homo sapiens

<400> 124

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Glu Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 125

<211> 122

<212> Protein

<213> Homo sapiens

<400> 125

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Lys Phe Arg Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Glu Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 126

<211> 107

<212> Protein

<213> Homo sapiens

<400> 126

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 127

<211> 122

<212> Protein

<213> Homo sapiens

<400> 127

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Lys Phe Arg Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Glu Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 128

<211> 107

<212> Protein

<213> Homo sapiens

<400> 128

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Asp Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Arg Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 129

<211> 122

<212> Protein

<213> Homo sapiens

<400> 129

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Lys Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 130

<211> 107

<212> Protein

<213> Homo sapiens

<400> 130

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Ile Lys Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 131

<211> 122

<212> Protein

<213> Homo sapiens

<400> 131

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Lys Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 132

<211> 107

<212> Protein

<213> Homo sapiens

<400> 132

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 133

<211> 122

<212> Protein

<213> Homo sapiens

<400> 133

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Thr Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 134

<211> 107

<212> Protein

<213> Homo sapiens

<400> 134

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 135

<211> 122

<212> Protein

<213> Homo sapiens

<400> 135

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Lys Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 136

<211> 107

<212> Protein

<213> Homo sapiens

<400> 136

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 137

<211> 122

<212> Protein

<213> Homo sapiens

<400> 137

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Lys Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 138

<211> 107

<212> Protein

<213> Homo sapiens

<400> 138

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 139

<211> 122

<212> Protein

<213> Homo sapiens

<400> 139

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Lys Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 140

<211> 107

<212> Protein

<213> Homo sapiens

<400> 140

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 141

<211> 122

<212> Protein

<213> Homo sapiens

<400> 141

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Gly Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 142

<211> 107

<212> Protein

<213> Homo sapiens

<400> 142

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 143

<211> 122

<212> Protein

<213> Homo sapiens

<400> 143

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 144

<211> 107

<212> Protein

<213> Homo sapiens

<400> 144

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Asn Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 145

<211> 122

<212> Protein

<213> Homo sapiens

<400> 145

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Lys Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 146

<211> 107

<212> Protein

<213> Homo sapiens

<400> 146

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Glu Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 147

<211> 122

<212> Protein

<213> Homo sapiens

<400> 147

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Lys Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ile Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 148

<211> 107

<212> Protein

<213> Homo sapiens

<400> 148

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 149

<211> 122

<212> Protein

<213> Homo sapiens

<400> 149

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 150

<211> 107

<212> Protein

<213> Homo sapiens

<400> 150

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 151

<211> 120

<212> Protein

<213> Homo sapiens

<400> 151

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Thr Ser

20 25 30

Trp Ile Val Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Phe Thr Ser Tyr Ser Pro Ser Phe

50 55 60

Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Leu His Tyr Tyr His Ser Glu Glu Phe Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 152

<211> 109

<212> Protein

<213> Homo sapiens

<400> 152

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Arg Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Phe Gly Ser Ser Arg

85 90 95

Leu Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 153

<211> 122

<212> Protein

<213> Homo sapiens

<400> 153

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 154

<211> 107

<212> Protein

<213> Homo sapiens

<400> 154

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Glu Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 155

<211> 120

<212> Protein

<213> Homo sapiens

<400> 155

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Thr Ser

20 25 30

Trp Ile Val Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Phe Thr Ser Tyr Ser Pro Ser Phe

50 55 60

Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Leu His Tyr Tyr His Ser Glu Glu Phe Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 156

<211> 109

<212> Protein

<213> Homo sapiens

<400> 156

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Arg Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Phe Gly Ser Ser Arg

85 90 95

Leu Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 157

<211> 120

<212> Protein

<213> Homo sapiens

<400> 157

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Thr Ser

20 25 30

Trp Ile Val Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Phe Thr Ser Tyr Ser Pro Ser Phe

50 55 60

Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Leu His Tyr Tyr His Ser Glu Glu Phe Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 158

<211> 109

<212> Protein

<213> Homo sapiens

<400> 158

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Arg Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Phe Gly Ser Ser Arg

85 90 95

Leu Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 159

<211> 119

<212> Protein

<213> Homo sapiens

<400> 159

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 160

<211> 112

<212> Protein

<213> Homo sapiens

<400> 160

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 161

<211> 123

<212> Protein

<213> Homo sapiens

<400> 161

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ala

20 25 30

His Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Arg Ile Lys Arg Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala

50 55 60

Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Thr Thr Asp Gly Ile Ala Val Ala Gly Pro Leu Arg Val Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 162

<211> 113

<212> Protein

<213> Homo sapiens

<400> 162

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Val Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Ile Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 163

<211> 119

<212> Protein

<213> Homo sapiens

<400> 163

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 164

<211> 112

<212> Protein

<213> Homo sapiens

<400> 164

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 165

<211> 123

<212> Protein

<213> Homo sapiens

<400> 165

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala

20 25 30

Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Arg Ile Lys Arg Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala

50 55 60

Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asp Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Thr Thr Asp Gly Ile Ala Val Ala Gly Pro Leu Arg Val Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 166

<211> 113

<212> Protein

<213> Homo sapiens

<400> 166

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Val Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Ile Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 167

<211> 119

<212> Protein

<213> Homo sapiens

<400> 167

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 168

<211> 112

<212> Protein

<213> Homo sapiens

<400> 168

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 169

<211> 123

<212> Protein

<213> Homo sapiens

<400> 169

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ala

20 25 30

Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala

50 55 60

Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Thr Thr Asp Gly Ile Ala Val Ala Gly Pro Leu Arg Val Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 170

<211> 113

<212> Protein

<213> Homo sapiens

<400> 170

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Gly Asn Tyr Leu Val Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Ile Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 171

<211> 122

<212> Protein

<213> Homo sapiens

<400> 171

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 172

<211> 107

<212> Protein

<213> Homo sapiens

<400> 172

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 173

<211> 123

<212> Protein

<213> Homo sapiens

<400> 173

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ala

20 25 30

Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala

50 55 60

Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Thr Thr Asp Gly Ile Ala Val Ala Gly Pro Leu Arg Val Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 174

<211> 113

<212> Protein

<213> Homo sapiens

<400> 174

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Gly Asn Tyr Leu Val Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Ile Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 175

<211> 119

<212> Protein

<213> Homo sapiens

<400> 175

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Arg Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ala Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 176

<211> 112

<212> Protein

<213> Homo sapiens

<400> 176

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Thr Trp Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Lys Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 177

<211> 122

<212> Protein

<213> Homo sapiens

<400> 177

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 178

<211> 107

<212> Protein

<213> Homo sapiens

<400> 178

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Thr Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 179

<211> 120

<212> Protein

<213> Homo sapiens

<400> 179

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Thr Ser

20 25 30

Trp Ile Val Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Phe Thr Ser Tyr Ser Pro Ser Phe

50 55 60

Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Leu His Tyr Tyr His Ser Glu Glu Phe Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 180

<211> 109

<212> Protein

<213> Homo sapiens

<400> 180

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Arg Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Phe Gly Ser Ser Arg

85 90 95

Leu Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 181

<211> 122

<212> Protein

<213> Homo sapiens

<400> 181

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Trp Arg Gly Asp Ile Ile Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Leu Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Val Lys Ser Tyr Gly Ser Gly Ser Phe Tyr Asn Ala Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 182

<211> 107

<212> Protein

<213> Homo sapiens

<400> 182

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ala Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile

35 40 45

Tyr Lys Ala Ser Arg Leu Asp Arg Gly Thr Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Leu Glu Tyr Ser Ser Tyr Ile Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 183

<211> 120

<212> Protein

<213> Homo sapiens

<400> 183

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Thr Ser

20 25 30

Trp Ile Val Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Phe Thr Ser Tyr Ser Pro Ser Phe

50 55 60

Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Leu His Tyr Tyr His Ser Glu Glu Phe Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 184

<211> 109

<212> Protein

<213> Homo sapiens

<400> 184

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Thr Arg Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Phe Gly Ser Ser Arg

85 90 95

Leu Phe Thr Phe Gly Glu Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 185

<211> 123

<212> Protein

<213> Homo sapiens

<400> 185

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala

20 25 30

Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Arg Ile Lys Arg Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala

50 55 60

Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asp Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Thr Thr Asp Gly Ile Ala Val Ala Gly Pro Leu Arg Val Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 186

<211> 113

<212> Protein

<213> Homo sapiens

<400> 186

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Val Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Ile Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 187

<211> 119

<212> Protein

<213> Homo sapiens

<400> 187

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Arg Pro Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asp Tyr Gly Pro Ser Phe

50 55 60

Glu Gly His Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Ile Arg Ser Gly Ala Phe Asp Ile Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 188

<211> 112

<212> Protein

<213> Homo sapiens

<400> 188

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Arg Leu Leu His Arg

20 25 30

Thr Gly Tyr Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Arg Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Val

85 90 95

Leu His Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<---

Claims (16)

1. A procoagulant bispecific antibody or antigen binding fragment thereof, comprising a first antigen binding region capable of binding to FIX described by SEQ ID NO: 1 and/or an activated form of FIXa thereof, and a second antigen binding region capable of binding to FX described by SEQ ID NO: 1. 2, and/or its activated form FXa, where a) the first antigen binding region contains a heavy chain variable domain that is at least 96% identical to the sequence of SEQ ID NO: 177, and a light chain variable domain that is at least 96% identical to the sequence of SEQ ID NO: 178, and where b) the second antigen binding region contains a heavy chain variable domain that is at least 96% identical to the sequence of SEQ ID NO: 21, and a light chain variable domain that is at least 96% identical to the sequence of SEQ ID NO: 22. 2. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein the first antigen-binding region is capable of binding an epitope containing amino acid residue R338 of FIX/FIXa, and the second antigen-binding site is capable of binding to the EGF-2 domain and/or the FX/FXa catalytic subunit. 3. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein the first antigen-binding region is capable of binding an epitope containing amino acid residues R338 and K341 of FIX/FIXa. 4. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein the first antigen-binding region is capable of binding an epitope containing amino acid residues L337, R338, S339, T340, K341 and T343 of FIX/FIXa. 5. A bispecific antibody or an antigen-binding fragment thereof according to claim 1, where the first antigen-binding region is capable of binding an epitope containing amino acid residues D332, R333, L337 and R338 FIX/FIXa, and the second antigen-binding region is capable of binding to the EGF-2 domain and/or catalytic subunit FX/FXa. 6. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein the first antigen-binding region is capable of binding an epitope containing amino acid residues K301, D332, R333, A334, T335, R338 and N346 of FIX/FIXa. 7. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein the first antigen-binding region is capable of binding an epitope containing amino acid residues R113, Y230, K420, D423, R424 and K427 from FX/FXa. 8. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein the second antigen-binding region is capable of binding an epitope containing one or more amino acid residues from H101, E103, R113, T116, L117, A118, T127, S227, E228, F229, Y230, E266, R287, L303, P304, E305, L419, K420, D423, R424, M426, K427 and T428 from FX/FXa. 9. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein the isotype of the antibody is IgG4. 10. The bispecific antibody or antigen-binding fragment thereof according to claim 1, wherein said bispecific antibody or antigen-binding fragment thereof is capable of stimulating the enzymatic activity of FIXa towards FX. 11. A pharmaceutical composition for the treatment of hemophilia A in the presence of inhibitors or hemophilia A in which there are no inhibitors, containing a bispecific antibody or an antigen-binding fragment according to any of the previous paragraphs and a pharmaceutically acceptable carrier. 12. A medicinal product for the treatment of hemophilia A in the presence of inhibitors or hemophilia A in which there are no inhibitors, containing a bispecific antibody or its antigen-binding fragment according to any one of paragraphs. 1-10.

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